Aryl ethene derivative and pharmaceutical composition containing same as active ingredient

ABSTRACT

The present invention relates to an aryl ethene derivative, for inhibiting an estrogen-related receptor gamma (ERRγ) activity, a prodrug of same, a solvate of same, a stereoisomer of same or pharmaceutically acceptable salts of same, and a pharmaceutical composition containing same as an active ingredient.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a U.S. National Phase of International PatentApplication Serial No. PCT/KR2016/010364 entitled “NOVEL ARYL ETHENEDERIVATIVE AND PHARMACEUTICAL COMPOSITION CONTAINING SAME AS ACTIVEINGREDIENT,” filed on Sep. 13, 2016. International Patent ApplicationSerial No. PCT/KR2016/010364 claims priority to Korean PatentApplication No. 10-2016-0080124, filed on Jun. 27, 2016. The entirecontents of each of the above-cited applications are hereby incorporatedby reference in their entirety for all purposes.

TECHNICAL FIELD

The present invention relates to an arylethene derivative inhibiting anactivity of an estrogen-related receptor gamma (hereinafter, referred toas ERRγ), or a prodrug, solvate, stereoisomer, or pharmaceuticallyacceptable salt thereof, and a pharmaceutical composition comprising thecompound as an active ingredient.

BACKGROUND ART

A hormone receptor which responds to the hormone is required forregulating development, growth or differentiation of cells throughchange in intracellular gene expression, and is largely classified intoa cell membrane receptor and a nuclear receptor. Among them, there is anincreasing interest in an orphan nuclear receptor which is the nuclearreceptor and of which the binding ligand has not been revealed.

Estrogen-related receptor (ERR) which is one of the orphan nuclearreceptors has three types which are ERRα, ERRβ, and ERRγ, and eachposition to be activated is different.

In particular, ERRγ shows an activity in spinal cords and a centralnervous system, and is a nuclear hormone receptor which is atranscriptional regulatory protein involved in glucose biosynthesis in aliver, and has an increased transcriptional activity for itself whenbound to a ligand, thereby helping gene expression related to glucosesynthesis. That is, ERRγ is directly involved in glucose metabolism.

In addition, ERRγ is a human nuclear receptor protein called NR3B3, andis encoded by a ESRRG gene. ERRγ functions as a constitutive activatorin transcription. ERRγ is a member of a nuclear hormone receptor familyof a steroid hormone receptor.

An ERRγ protein is known as a main modulator of various genes related tofatty acid oxidation and mitochondria biogenesis in a myocardium, andalso known to be involved in glucose production in a liver.

Meanwhile, diabetic retinopathy is a disease developed by occurrence ofcirculatory failure in a retina which is specific to diabetic patients,and belongs to one of the three major microvascular complications ofdiabetes together with diabetic neuropathy and diabetic nephropathy.Occurrence of diabetic retinopathy is related to a disease period duringwhich a patient suffers from diabetes, and in the case of the diabetesdiagnosed before the age of 30 corresponding to type 1, the diabeticretinopathy occurs in 17% when the disease period is 5 years or less,and in 98% when the disease period is 15 years or more, and among them,worsening proliferative diabetic retinopathy occurs in about 1% when thedisease period is 10 years or less, and in 67% when the disease periodis 35 years or more. In the case of type 2 diabetes, it is known thatthe diabetic retinopathy occurs in 29% when the disease period is 5years or less, and in 78% when the disease period is 15 years or more,and the proliferative diabetic retinopathy occurs in 2% when the diseaseperiod is 5 years or less, and in 16% when the disease period is 15years or more. In a diabetic patient's retina, it is known that vascularchange in a capillary such as hypertrophy of a retinal capillarybasement membrane, loss of perivascular cells, and occurrence ofmicroaneurysm occurs, and as time passes, retinal neovascularizationsubsequent to a wide range of capillary nonperfusion may also occur.This diabetic retinopathy is a kind of diabetes complications, but oncedevelops, the progression thereof is difficult to be prevented byglycemic control, and a treatment method specific to retinopathy isdemanded.

It has been reported from a recent study that a low molecular organiccompound known as GS K5182 which is(Z)-4-(1-(4-(2-(dimethylamino)ethoxy)phenyl)-5-hydroxy-2-phenylpent-1-en-1-yl)phenolfunctions as a ligand in ERRγ to inhibit the ERRγ activity, therebyshowing an anti-diabetes effect such as relieving hyperglycemia andinsulin resistance, and a treatment effect of retinopathy.

Development of a new material which significantly inhibits atranscriptional activity of ERRγ as compared with previously reportedGSK5182 is demanded.

Meanwhile, anaplastic thyroid cancer (ATC) is one of the most aggressiveand deadly cancers known to develop in humans. ATC rapidly metastasizesfrom a thyroid gland to lungs, bones, focal lymph nodes, and the brain.This is in contrast with the nature of well-differentiated benignthyroid cancer which explains most of the thyroid cancer, and thus,treatment of ATC which is surgery, a radiation therapy, and achemotherapy alone or in combination thereof has not exhibited an effecton patient survival. As a result, development of a novel treatmentmethod is urgently demanded.

A sodium iodide symporter (NIS) is a plasma membrane glycoprotein whichmediates intracellular active inflow of iodine. In the treatment ofthyroid cancer, endogenous NIS accepts a wide range of application of aradioiodine therapy in a clinical situation, which is known as aneffective treatment method to remove malignant cells with minimal sideeffects over the years. Low-differentiated cancer cells including ATCcells tend to represent gradual dedifferentiation leading to a decreasein a NIS level. This prevents ATC cells from accumulating iodine in thecells with a high concentration, and accordingly, causes cell resistanceto the radioiodine therapy, leading to a poor prognosis. Therefore,there has been many attempts to recover an NIS function from ATC cells,using several methods such as epigenetic regulation using gene transfer,an epigenome-altering drug, and the like, however, no satisfactoryresult has been obtained so far.

The biological effect of ERRγ has been extensively studied in variousdisease models (type 2 diabetes mellitus, alcohol-derived oxidativestress, microbial infection by liver damage and gluconeogenesis of thedamaged liver, some metabolic diseases such as liver insulin signalingand iron metabolism), however, the role of ERRγ for the NIS function inATC has not been clearly studied so far. It has been reported from arecent study that a low molecular organic compound known as GSK5182which is(Z)-4-(1-(4-(2-(dimethylamino)ethoxy)phenyl)-5-hydroxy-2-phenylpent-1-en-1-yl)phenolfunctions as a ligand in ERRγ to inhibit the ERRγ activity, therebyimproving the function of NIS to increase an ATC intracellularradioiodine uptake and finally exhibit an effect of increasingradioiodine treatment. However, when GSK5182 was administered to an ATCmouse tumor model, a radioiodine uptake in the tumor was not increased.Accordingly, development of a new material which may specifically andsignificantly inhibit ERRγ transcriptional activity as compared withGSK5182, and as a result, cause a radioactive isotope uptake increasefrom a cellular level to an animal level is demanded.

DISCLOSURE Technical Problem

Thus, the inventors of the present invention found that by introducing aspecific substituent to an arylethene derivative, an activity to inhibitERRγ is better as compared with the conventionally reported activity ofGSK5182, and at the same time, drug stability, a pharmacologicalactivity, and toxicity were improved, thereby completing the presentinvention.

An object of the present invention is to provide a novel arylethenederivative which may effectively inhibit an ERRγ activity, or a prodrug,solvate, stereoisomer, or pharmaceutically acceptable salt thereof.

Another object of the present invention is to provide a pharmaceuticalcomposition for preventing or treating ERRγ-mediated diseases,comprising the arylethene derivative, or the prodrug, solvate,stereoisomer, or pharmaceutically acceptable salt thereof as an activeingredient.

Another object of the present invention is to provide a pharmaceuticalcomposition for preventing or treating retinopathy, comprising thearylethene derivative, or the prodrug, solvate, stereoisomer, orpharmaceutically acceptable salt thereof as an active ingredient.

Another object of the present invention is to provide a pharmaceuticalcomposition for treating thyroid cancer, comprising the arylethenederivative, or the prodrug, solvate, stereoisomer, or pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable carrier, andbeing used in combination of radioactive iodine.

Another object of the present invention is to provide a kit for treatingthyroid cancer, comprising the arylethene derivative, or the prodrug,solvate, stereoisomer, or pharmaceutically acceptable salt thereof, andradioiodine.

Technical Solution

In one general aspect, an arylethene derivative represented by thefollowing Chemical Formula 1, as a novel compound which may effectivelyinhibit an activity of ERRγ, or a prodrug, solvate, stereoisomer, orpharmaceutically acceptable salt thereof:

wherein

L is (C6-C20)arylene, (C3-C20)heteroarylene, or (C3-C20) fusedheterocycle;

R¹ is (C3-C20)heterocycloalkyl, (C3-C20)heteroaryl, —O—(CH₂)_(m)—R¹¹,—(CH₂)_(m)—R¹², —NH—(CH₂)_(m)—R¹³, —NHCO—(CH₂)_(n)—R¹⁴, or—SiR¹⁶R¹⁷—(CH₂)_(m)—R¹⁵;

R¹¹ to R¹⁵ are independently of one another (C3-C20)heterocycloalkyl;

R¹⁶ and R¹⁷ are independently of each other (C1-C20)alkyl;

m is an integer of 1 to 3;

n is an integer of 0 or 1;

Ar is (C6-C20)aryl or (C3-C20)heteroaryl, in which the aryl orheteroaryl of Ar may be further substituted by one or more selected fromthe group consisting of hydroxy, halogen, (C1-C20)alkyl,halo(C1-C20)alkyl, (C1-C20)alkoxy, nitro, cyano, —NR²¹R²²,(C1-C20)alkylcarbonyloxy, (C1-C20)alkylcarbonylamino, guanidino,—SO₂—R²³, and —OSO₂—R²⁴;

R²¹ and R²² are independently of each other hydrogen,(C1-C10)alkylsulfonyl, or (C6-C20)cycloalkylsulfonyl;

R²³ and R²⁴ are independently of each other (C1-C20)alkyl,halo(C1-C20)alkyl, or (C3-C20)cycloalkyl;

R² is hydroxy, halogen, (C1-C20)alkylcarbonyloxy, or(C1-C20)alkylsulfonyloxy;

the heterocycloalkyl or heteroaryl of R¹ and the heterocycloalkyl of R¹¹to R¹⁵ may be further substituted by one or more selected from the groupconsisting of (C1-C20)alkyl, (C3-C20)cycloalkyl, (C2-C20)alkenyl,amidino, (C1-C20)alkoxycarbonyl, hydroxy, hydroxy(C1-C20)alkyl, anddi(C1-C20)alkylamino(C1-C20)alkyl; and

the heterocycloalkyl and heteroaryl contains one or more heteroatomsselected from the group consisting of N, O and S, and theheterocycloalkyl is a saturated or unsaturated mono-, bi-, or spirocyclehaving a carbon atom or nitrogen atom in a ring as a binding site.

In another general aspect, a pharmaceutical composition for preventingor treating ERRγ-mediated diseases includes: the arylethene derivative,or a prodrug, solvate, stereoisomer, or pharmaceutically acceptable saltthereof as an active ingredient, by confirming an excellent ERRγinhibitory activity of the arylethene derivative represented by ChemicalFormula 1.

In another general aspect, a pharmaceutical composition for preventingor treating retinopathy includes: the arylethene derivative of ChemicalFormula 1 which may effectively inhibit an ERRγ activity, or a prodrug,solvate, stereoisomer, or pharmaceutically acceptable salt thereof as anactive ingredient.

In another general aspect, a pharmaceutical composition for treatingthyroid cancer includes: the arylethene derivative of Chemical Formula 1which may specifically and significantly inhibit an ERRγ transcriptionalactivity, or a prodrug, solvate, stereoisomer, or pharmaceuticallyacceptable salt thereof, and a pharmaceutically acceptable carrier, andis used in combination of radioactive iodine.

In still another general aspect, a kit for treating thyroid cancerincludes: the arylethene derivative of Chemical Formula 1 which mayspecifically and significantly inhibit an ERRγ transcriptional activity,or a prodrug, solvate, stereoisomer, or pharmaceutically acceptable saltthereof, and radioactive iodine.

Advantageous Effects

The arylethene derivative of the present invention is a novel compound,and exhibits very high inhibitory activity to ERRγ as compared with aconventional GSK5182 compound, and at the same time, shows an effect ofimproved drug stability, pharmacological activity and toxicity. Thus,the arylethene derivative may be useful as efficient prophylactic agentand therapeutic agent for diseases mediated by ERRγ, in particular,metabolic diseases such as obesity, diabetes, hyperlipidemia, fattyliver, or atherosclerosis, as well as retinopathy, without side effects.

In addition, the arylethene derivative of the present invention mayspecifically and significantly inhibit ERRγ transcriptional activity ascompared with GSK5182, and as a result, cause a radioactive isotopeuptake increase from a cellular level to an animal level. Accordingly,the arylethene derivative of the present invention may significantlyincrease a treatment effect of radioactive iodine therapy for treatingcancer, and when administered to cancer cells, may effectively producecancer cells having an improved sodium iodide symporter (NIS) function,thereby having an excellent effect of being more easily applied torelated research and clinical practice for treating anaplastic thyroidcancer.

DESCRIPTION OF DRAWINGS

FIGS. 1 to 3 illustrate an effect of compound 18a for a radioactiveiodine uptake in anaplastic thyroid cancer cells.

FIGS. 4 and 5 illustrate an effect of compound 18a for regulatingendogenous ERRγ and NIS mRNA expression in anaplastic thyroid cancercells.

FIGS. 6 and 7 illustrate an effect of compound 18a for regulatingendogenous ERRγ protein expression in anaplastic thyroid cancer cells.

FIGS. 8 and 9 illustrate a compound 18a-derived MAP kinase activity inanaplastic thyroid cancer cells.

FIGS. 10 and 11 illustrate a degree of iodine uptake inhibition incompound 18a-treated anaplastic thyroid cancer cells, by PD98059 orU0126.

FIGS. 12 and 13 illustrate a degree of inversion of activated MAK kinasesignaling, by PD98059 or U0126.

FIGS. 14 and 15 illustrate an increase aspect of an amount ofmembrane-localized NIS protein in anaplastic thyroid cancer cells bycompound 18a.

FIGS. 16 and 17 illustrate results showing increased cytotoxicity ofincreased ¹³¹I after treating anaplastic thyroid cancer cells withcompound 18a.

FIGS. 18 to 22 illustrate an effect of compound 18a for a radioactiveiodine uptake by administrating compound 18a in an ATC tumor model.

BEST MODE

Hereinafter, the present invention will be described in detail.Technical terms and scientific terms used in the present specificationhave the general meaning understood by those skilled in the art to whichthe present invention pertains unless otherwise defined, and adescription for the known function and configuration obscuring thepresent invention will be omitted in the following description.

The present invention provides an arylethene derivative represented bythe following Chemical Formula 1, or a prodrug, solvate, stereoisomer,or pharmaceutically acceptable salt thereof:

wherein

L is (C6-C20)arylene, (C3-C20)heteroarylene, or (C3-C20) fusedheterocycle;

R¹ is (C3-C20)heterocycloalkyl, (C3-C20)heteroaryl, —O—(CH₂)_(m)—R¹¹,—(CH₂)_(m)—R¹², —NH—(CH₂)_(m)—R¹³, —NHCO—(CH₂)_(n)—R¹⁴, or—SiR¹⁶R¹⁷—(CH₂)_(m)—R¹⁵;

R¹¹ to R¹⁵ are independently of one another (C3-C20)heterocycloalkyl;

R¹⁶ and R¹⁷ are independently of each other (C1-C20)alkyl;

m is an integer of 1 to 3;

n is an integer of 0 or 1;

Ar is (C6-C20)aryl or (C3-C20)heteroaryl, in which the aryl orheteroaryl of Ar may be further substituted by one or more selected fromthe group consisting of hydroxy, halogen, (C1-C20)alkyl,halo(C1-C20)alkyl, (C1-C20)alkoxy, nitro, cyano, —NR²¹R²²,(C1-C20)alkylcarbonyloxy, (C1-C20)alkylcarbonylamino, guanidino,—SO₂—R²³, and —OSO₂—R²⁴;

R²¹ and R²² are independently of each other hydrogen,(C1-C10)alkylsulfonyl, or (C6-C20)cycloalkylsulfonyl;

R²³ and R²⁴ are independently of each other (C1-C20)alkyl,halo(C1-C20)alkyl, or (C3-C20)cycloalkyl;

R² is hydroxy, halogen, (C1-C20)alkylcarbonyloxy, or(C1-C20)alkylsulfonyloxy;

the heterocycloalkyl or heteroaryl of R¹ and the heterocycloalkyl of R¹¹to R¹⁵ may be further substituted by one or more selected from the groupconsisting of (C1-C20)alkyl, (C3-C20)cycloalkyl, (C2-C20)alkenyl,amidino, (C1-C20)alkoxycarbonyl, hydroxy, hydroxy(C1-C20)alkyl, anddi(C1-C20)alkylamino(C1-C20)alkyl; and

the heterocycloalkyl and heteroaryl contains one or more heteroatomsselected from the group consisting of N, O and S, and theheterocycloalkyl is a saturated or unsaturated mono-, bi-, or spirocyclehaving a carbon atom or nitrogen atom in a ring as a binding site.

The arylethene derivative of Chemical Formula 1 according to the presentinvention which is a novel compound, has a very high inhibitory activityto ERRγ, and thus, is useful as a therapeutic agent and a prophylacticagent of ERRγ-mediated diseases, in particular, metabolic diseases suchas obesity, diabetes, hyperlipidemia, fatty liver or arteriosclerosis,and also may be used as an active ingredient for preventing or treatingretinopathy.

In addition, the arylethene derivative of Chemical Formula 1 accordingto the present invention regulates expression of endogenous ERRγ proteinto regulate mitogen-activated protein (MAP) kinase, and improves asodium iodide symporter (NIS) function to increase membrane-localizedNIS, thereby increasing a radioactive iodine uptake when treatingthyroid cancer.

The term of the present invention, “alkyl” refers to a monovalentstraight-chain or branched-chain saturated hydrocarbon radicalconsisting of only carbon and hydrogen atoms, and an example of thealkyl radical includes methyl, ethyl, propyl, isopropyl, butyl,isobutyl, t-butyl, pentyl, hexyl, octyl, nonyl, or the like, but notlimited thereto.

The term of the present invention, “aryl” refers to a monovalent organicradical of an aromatic ring derived from aromatic hydrocarbon by removalof one hydrogen, including a single- or fused ring system containingappropriately 4 to 7, preferably 5 or 6 ring atoms in each ring, andeven a form in which a plurality of aryls are linked by a single bond. Aspecific example thereof includes phenyl, naphthyl, biphenyl, anthryl,indenyl, fluorenyl, or the like, but not limited thereto.

The term of the present invention, “heteroaryl” refers to a monovalentradical of a heteroaromatic ring which is an aryl group containing 1 to4 heteroatoms selected from the group consisting of N, O, and S as anaromatic ring backbone atom, and carbons as remaining aromatic ringbackbone atoms, and is a 5- or 6-membered monocyclic heteroaryl and apolycyclic heteroaryl fused with one or more benzene rings, which may bepartially saturated. In addition, the heteroaryl in the presentinvention also includes a form in which one or more heteroaryls arelinked by a single bond. An example of the heteroaryl group includespyrrolyl, pyrazolyl, quinolyl, isoquinolyl, pyridyl, pyrimidinyl,oxazolyl, thiazolyl, thiadiazolyl, triazolyl, imidazolyl,benzimidazolyl, isoxazolyl, benzisoxazolyl, thiophenyl, benzothiophenyl,furyl, benzofuryl, or the like, but not limited thereto.

The term of the present invention, “arylene” and “heteroarylene” referto divalent radicals of aromatic ring and heteroaromatic ring.

The term of the present invention, “fused heterocycle” refers to adivalent radical of a fused ring in which a non-aromatic heterocyclecontaining 1 to 4 heteroatoms selected from the group consisting of N,O, and S, and an aromatic ring are fused, and has a carbon atom or anitrogen atom in the fused heterocycle as a bonding site. An example ofthe fused heterocycle includes indoline, dihydrobenzofuran,dihydrobenzothiophene, or the like, but not limited thereto.

The term of the present invention, “heterocycloalkyl” is a monovalentradical of a non-aromatic heterocycle containing 1 to 4 heteroatomsselected from the group consisting of N, O, and S, and the non-aromaticheterocycle includes a saturated or unsaturated monocycle, polycycle orspirocycle form, and may be bonded via a heteroatom or a carbon atom. Anexample of the heterocycloalkyl radical may include monovalent radicalsof non-aromatic heterocycles such as aziridine, pyrrolidine, azetidine,piperidine, tetrahydropyridine, piperazine, morpholine, thiomorpholine,3-azabicyclo[3.1.0]hexane, octahydropyrrolo[3,4-c]pyrrole,2,7-diazispiro[4.4]nonane, 2-azaspiro[4.4]nonane, or the like.

The term of the present invention, “halo” or “halogen” refers tofluorine, chlorine, bromine or iodine atom.

The term or the present invention, “haloalkyl” refers to alkylsubstituted by one or more halogens, and an example thereof may includetrifluoromethyl, or the like.

The term of the present invention, “alkenyl” is a monovalent radical ofa straight chain or branched chain unsaturated hydrocarbon including oneor more double bonds between two or more carbon atoms, and specificallyincludes ethenyl, propenyl, prop-1-en-2-yl, 1-butenyl, 2-butenyl,isobutylenyl, 1-pentenyl, 2-pentenyl, 3-methyl-1-butenyl,2-methyl-2-butenyl, 2,3-dimethyl-2-butenyl, or the like, but not limitedthereto.

The term of the present invention, “alkoxy” refers to an —O-alkylradical, wherein the alkyl is as described above. An example of thealkoxy radical includes methoxy, ethoxy, isopropoxy, butoxy, isobutoxy,t-butoxy, or the like, but not limited thereto.

The term of the present invention, “alkylcarbonyloxy” refers to an—OC(═O)alkyl radical, wherein the alkyl is as described above. Anexample of the alkylcarbonyloxy radical includes methylcarbonyloxy,ethylcarbonyloxy, isopropylcarbonyloxy, propylcarbonyloxy,butylcarbonyloxy, isobutylcarbonyloxy, t-butylcarbonyloxy, or the like,but not limited thereto.

The term of the present invention, “alkylcarbonylamino” refers to a—NHC(═O)alkyl radical, wherein the alkyl is as described above. Anexample of the alkylcarbonylamino radical includes methylcarbonylamino,ethylcarbonylamino, isopropylcarbonylamino, propylcarbonylamino,butylcarbonylamino, isobutylcarbonylamino, t-butylcarbonylamino, or thelike, but not limited thereto.

The term of the present invention, “alkoxycarbonyl” refers to a—C(═O)alkoxy radical, wherein the alkoxy is as described above. Anexample of the alkoxycarbonyl radical includes methoxycarbonyl,ethoxycarbonyl, isopropoxycarbonyl, propoxycarbonyl, butoxycarbonyl,isobutoxycarbonyl, t-butoxycarbonyl, or the like, but not limitedthereto.

The term of the present invention, “cycloalkyl” refers to a monovalentsaturated carbocyclic radical composed of one or more rings. An exampleof the cycloalkyl radical includes cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, or the like, but not limited thereto.

The term of the present invention, “alkylsulfonyl” refers to a—SO₂-alkyl radical, wherein the alkyl is as described above. An exampleof the alkylsulfonyl radical includes methylsulfonyl, ethylsulfonyl, orthe like, but not limited thereto.

The term of the present invention, “cycloalkylsulfonyl” refers to a—SO₂-cycloalkyl radical, wherein the cycloalkyl is as described above.An example of the cycloalkylsulfonyl radical includescyclopropylsulfonyl, cyclohexylsulfonyl, or the like, but not limitedthereto.

The term of the present invention, “alkylsulfonyloxy” refers to a—OSO₂-alkyl radical, wherein the alkyl is as described above. An exampleof the alkylsulfonyloxyl radical includes methylsulfonyloxy,ethylsulfonyloxy, or the like, but not limited thereto.

The term or the present invention, “hydroxyalkyl” refers to alkylsubstituted by one or more hydroxys, and an example thereof may includehydroxymethyl or the like.

In the arylethene derivative according to an exemplary embodiment of thepresent invention, the arylethene derivative may be represented by thefollowing Chemical Formulae 2 to 5:

wherein

denotes a single bond or a double bond; and R¹, Ar and R² are as definedin the above Chemical Formula 1.

In the arylethene derivative according to an exemplary embodiment of thepresent invention, R¹ is (C3-C10)heterocycloalkyl, (C3-C10)heteroaryl,—O—(CH)_(m)—R¹¹, —(CH₂)_(m)—R¹², —NH—(CH₂)_(m)—R¹³, —NHCO—(CH₂)_(n)—R¹⁴,or —SiR¹⁶R¹⁷—(CH₂)_(m)—R¹⁵; R¹¹ to R¹⁵ are independently of one another(C3-C10)heterocycloalkyl; R¹⁶ and R¹⁷ are independently of each other(C1-C10)alkyl; m is an integer of 1 to 3; n is an integer of 0 or 1; Aris (C6-C12)aryl or (C3-C12)heteroaryl, in which the aryl or heteroarylof Ar may be further substituted by one or more selected from the groupconsisting of hydroxy, halogen, (C1-C10)alkyl, halo(C1-C10)alkyl,(C1-C10)alkoxy, nitro, cyano, amino, (C1-C10)alkylsulfonylamino,(C3-C10)cycloalkylsulfonylamino, di((C1-C10)alkylsulfonyl)amino,(C1-C10)alkylcarbonyloxy, (C1-C10)alkylcarbonylamino, guanidino,(C1-C10)alkylsulfonyl, (C1-C10)alkylsulfonyloxy,halo(C1-C10)alkylsulfonyloxy, and (C3-C10)cycloalkylsulfonyloxy; R² ishydroxy, fluoro, (C1-C10)alkylcarbonyloxy, or (C1-C10)alkylsulfonyloxy;and the heterocycloalkyl or heteroaryl of R¹ and the heterocycloalkyl ofR¹¹ to R¹⁵ may be further substituted by one or more selected from thegroup consisting of (C1-C10)alkyl, (C3-C10)cycloalkyl, (C2-C10)alkenyl,amidino, (C1-C10)alkoxycarbonyl, hydroxy(C1-C10)alkyl, anddi(C1-C10)alkylamino(C1-C10)alkyl.

In the arylethene derivative according to an exemplary embodiment of thepresent invention, it is preferred that R¹ is (C3-C10)heterocycloalkylor —O—(CH₂)_(m)—R¹¹; R¹¹ is (C3-C10)heterocycloalkyl; m is an integer of1 to 3; and the heterocycloalkyl of R¹ and R¹¹ may be furthersubstituted by one or more selected from the group consisting of(C1-C10)alkyl, (C3-C10)cycloalkyl, (C2-C10)alkenyl, amidino,(C1-C10)alkoxycarbonyl, hydroxy(C1-C10)alkyl, anddi(C1-C10)alkylamino(C1-C10)alkyl.

In the arylethene derivative according to an exemplary embodiment of thepresent invention, it is more preferred that heterocycloalkyl of the R¹and R¹¹ to R¹⁵ may be independently of each other selected from thefollowing structures:

wherein R³¹ and R³² are independently of each other hydrogen,(C1-C10)alkyl, (C3-C10)cycloalkyl, (C2-C10)alkenyl, amidino,(C1-C10)alkoxycarbonyl, hydroxy(C1-C10)alkyl, ordi(C1-C10)alkylamino(C1-C10)alkyl; and L is O or S.

In the arylethene derivative according to an exemplary embodiment of thepresent invention, the arylethene derivative may be more preferablyrepresented by the following Chemical Formula 6:

wherein

R¹ is (C3-C10)heterocycloalkyl or —O—(CH₂)_(m)—R¹¹;

R¹¹ is (C3-C10)heterocycloalkyl;

m is an integer of 1 to 3;

the heterocycloalkyl of R¹ and R¹¹ may be further substituted by one ormore selected from the group consisting of (C1-C10)alkyl,(C3-C10)cycloalkyl, (C2-C10)alkenyl, amidino, (C1-C10)alkoxycarbonyl,hydroxy(C1-C10)alkyl, and di(C1-C20)alkylamino(C1-C20)alkyl;

Ar is (C6-C12)aryl or (C3-C12)heteroaryl, in which the aryl orheteroaryl of Ar may be further substituted by one or more selected fromthe group consisting of hydroxy, halogen, (C1-C10)alkyl,halo(C1-C10)alkyl, (C1-C10)alkoxy, nitro, cyano, amino,(C1-C10)alkylsulfonylamino, (C3-C10)cycloalkylsulfonylamino,di((C1-C10)alkylsulfonyl)amino, (C1-C10)alkylcarbonyloxy,(C1-C10)alkylcarbonylamino, guanidino, (C1-C10)alkylsulfonyl,(C1-C10)alkylsulfonyloxy, halo(C1-C10)alkylsulfonyloxy, and(C3-C10)cycloalkylsulfonyloxy; and

R² is hydroxy, fluoro, (C1-C10)alkylcarbonyloxy, or(C1-C10)alkylsulfonyloxy.

In the arylethene derivative according to an exemplary embodiment of thepresent invention, R¹ and R¹¹ may be independently of each otherheterocycloalkyl selected from the following structures:

wherein R³¹ and R³² are independently of each other hydrogen,(C1-C10)alkyl, (C3-C10)cycloalkyl, (C2-C10)alkenyl, amidino,(C1-C10)alkoxycarbonyl, hydroxy(C1-C10)alkyl, ordi(C1-C10)alkylamino(C1-C10)alkyl; and L is O or S.

In the arylethene derivative according to an exemplary embodiment of thepresent invention, Ar is (C6-C20)aryl, in which the aryl of Ar may befurther substituted by one or more selected from the group consisting ofhydroxy, halogen, (C1-C10)alkyl, halo(C1-C10)alkyl, (C1-C10)alkoxy,nitro, cyano, amino, (C1-C10)alkylsulfonylamino,(C3-C10)cycloalkylsulfonylamino, di((C1-C10)alkylsulfonyl)amino,(C1-C10)alkylcarbonyloxy, (C1-C10)alkylcarbonylamino, guanidino,(C1-C10)alkylsulfonyl, (C1-C10)alkylsulfonyloxy,halo(C1-C10)alkylsulfonyloxy, and (C3-C10)cycloalkylsulfonyloxy.

In the arylethene derivative according to an exemplary embodiment of thepresent invention, R² may be hydroxy.

In the arylethene derivative according to an exemplary embodiment of thepresent invention, R² may be hydroxy, and R¹ may be heterocycloalkylselected from the following structures:

wherein R³¹ and R³² are independently of each other hydrogen,(C1-C10)alkyl, (C3-C10)cycloalkyl, (C2-C10)alkenyl, amidino,(C1-C10)alkoxycarbonyl, hydroxy(C1-C10)alkyl, ordi(C1-C10)alkylamino(C1-C10)alkyl; and L is O or S.

In the arylethene derivative according to an exemplary embodiment of thepresent invention, it is more preferred that R² is hydroxy and R¹ is—O—(CH₂)_(m)—R¹¹; m is an integer of 1 or 2; and R¹¹ is heterocycloalkylselected from the following structures:

wherein R³¹ and R³² are independently of each other hydrogen,(C1-C10)alkyl, (C-C10)alkoxycarbonyl, or hydroxy(C1-C10)alkyl; and L isO or S.

In the arylethene derivative according to an exemplary embodiment of thepresent invention, it is more preferred that Ar is (C6-C12)aryl, inwhich the aryl of Ar may be further substituted by one or more selectedfrom the group consisting of hydroxy, halogen, (C1-C10)alkyl,halo(C1-C10)alkyl, (C1-C10)alkoxy, nitro, cyano, amino,(C1-C10)alkylsulfonylamino, (C3-C10)cycloalkylsulfonylamino,di((C1-C10)alkylsulfonyl)amino, (C1-C10)alkylcarbonyloxy,(C1-C10)alkylcarbonylamino, guanidino, (C1-C10)alkylsulfonyl,(C1-C10)alkylsulfonyloxy, halo(C1-C10)alkylsulfonyloxy, and(C3-C10)cycloalkylsulfonyloxy; R² is hydroxy; and R¹ is heterocycloalkylselected from the following structures:

wherein R³¹ and R³² are independently of each other hydrogen,(C1-C10)alkyl, (C3-C10)cycloalkyl, (C2-C10)alkenyl, amidino,(C1-C10)alkoxycarbonyl, hydroxy(C1-C10)alkyl, ordi(C1-C10)alkylamino(C1-C10)alkyl; and L is O or S.

In the arylethene derivative according to an exemplary embodiment of thepresent invention, it is still more preferred that Ar is (C6-C12)aryl,in which the aryl of Ar may be further substituted by one or moreselected from the group consisting of hydroxy, halogen, (C1-C10)alkyl,halo(C1-C10)alkyl, (C1-C10)alkoxy, nitro, cyano, amino,(C1-C10)alkylsulfonylamino, (C3-C10)cycloalkylsulfonylamino,di((C1-C10)alkylsulfonyl)amino, (C1-C10)alkylcarbonyloxy,(C1-C10)alkylcarbonylamino, guanidino, (C1-C10)alkylsulfonyl,(C1-C10)alkylsulfonyloxy, halo(C1-C10)alkylsulfonyloxy, and(C3-C10)cycloalkylsulfonyloxy; R² is hydroxy; R¹ is —O—(CH₂)_(m)—R¹¹, mis an integer of 1 or 2, R¹¹ is heterocycloalkyl selected from thefollowing structures:

wherein R³¹ and R³² are independently of each other hydrogen,(C1-C20)alkyl, (C-C10)alkoxycarbonyl, or hydroxy(C1-C10)alkyl; and L isO or S.

In the arylethene derivative according to an exemplary embodiment of thepresent invention, the arylethene derivative may be specificallyselected from the following structure, but not limited thereto:

In the arylethene derivative according to an exemplary embodiment of thepresent invention, the arylethene derivative may be preferably selectedfrom the following structures:

In the arylethene derivative according to an exemplary embodiment of thepresent invention, the arylethene derivative may be more preferablyselected from the following structures:

In the arylethene derivative according to an exemplary embodiment of thepresent invention, the arylethene derivative may be still morepreferably selected from the following structures:

Since the arylethene derivative according to the present invention maybe used in the form of a prodrug, solvate, and pharmaceuticallyacceptable salt thereof for increasing in vivo absorption or increasingsolubility, the prodrug, the solvate, and the pharmaceuticallyacceptable salt also fall within the scope of the present invention. Inaddition, since the arylethene derivative has a chiral carbon, thestereoisomer thereof exists, and the stereoisomer also falls within thescope of the present invention.

The arylethene derivative according to the present invention may beprepared by various methods known in the art depending on the kinds ofsubstituents, and as an example thereof, the following Reaction Formulae1 to 21 are illustrated, and the following preparation methods do notlimit a method of preparing the arylethene derivative of the presentinvention. The specific details will be described in the followingExamples 1 to 121. The preparation methods presented in the followingReaction Formulae 1 to 21 are only illustrative, and it is apparent to aperson skilled in the art that the preparation methods may be easilymodified by a person skilled in the art depending on certainsubstituents.

In addition, the present invention provides an ERRγ inhibitorcomposition comprising the arylethene derivative of Chemical Formula 1,or the prodrug, solvate, stereoisomer, or pharmaceutically acceptablesalt thereof as an active ingredient.

In addition, the present invention provides a pharmaceutical compositionfor preventing or treating an ERRγ-mediated disease, comprising thearylethene derivative of Chemical Formula 1, or the prodrug, solvate,stereoisomer, or pharmaceutically acceptable salt thereof as an activeingredient, and further a pharmaceutically acceptable carrier.

As described above, since the arylethene derivative of Chemical Formula1, or the prodrug, solvate, stereoisomer, or pharmaceutically acceptablesalt thereof exhibits a high inhibitory activity to ERRγ, apharmaceutically acceptable composition comprising them as an activeingredient may be useful for treating or preventing ERRγ-mediateddiseases, for example, metabolic diseases such as obesity, diabetes,hyperlipidemia, fatty liver, or atherosclerosis.

In another general aspect, a pharmaceutical composition for preventingor treating retinopathy includes: the arylethene derivative of ChemicalFormula 1 which may effectively inhibit an ERRγ activity, or a prodrug,solvate, stereoisomer, or pharmaceutically acceptable salt thereof as anactive ingredient.

The “retinopathy” is a disease caused by chronic or acute damage to aretina of an eye. The retinopathy may involve ongoing inflammation andvascular remodeling. In addition, retinopathy also appears as visualmanifestation of a systemic disease such as diabetes or hypertension.The kind of retinopathy includes diabetic retinopathy, retinopathy ofprematurity (ROP), or the like.

Here, the diabetic retinopathy refers to an eye complication in whichdecreased visual acuity occurs due to a disorder following a peripheralcirculatory disorder caused by diabetes which is a systemic disease.Diabetic retinopathy has no symptoms at the beginning, but as macularinvasion occurs, decreased visual acuity appears. Diabetic retinopathyinvolves various pathological features such as microaneurysm,phlebectasia, retinal hemorrhage, retinal infarction, macular edema,neovascularization, vitreous hemorrhage, traction membrane, or the like,and when these phenomena are observed as ocular fundus symptoms,diabetic retinopathy is diagnosed. The diabetic retinopathy is a diseasecaused by a complex combination of various symptoms as described above,and it is unclear whether the disease is treated when one of thesesymptoms is alleviated.

In addition, retinopathy of prematurity is proliferative retinopathywhich may occur in premature babies, in particular low birth weightinfants. When a premature baby whose retinal blood vessels are notcompletely formed at birth has failure in angiogenesis process afterbirth, abnormal fibrovascular proliferation occurs at a border of anangiogenic site and a non-angiogenic site of a retina, whereby theretina is detached, eventually leading to blindness.

The arylethene derivative according to the present invention may be usedin the form of a pharmaceutically acceptable salt, and thepharmaceutically acceptable salt may be prepared by a conventionalmethod in the art, and may include for example, a salt with an inorganicacid such as a hydrochloric acid, a bromic acid, a sulfuric acid, sodiumhydrogen sulfate, a phosphoric acid, a nitric acid, or a carbonic acid,a salt with an organic acid such as a formic acid, an acetic acid, atrifluoroacetic acid, a propionic acid, an oxalic acid, a succinic acid,a benzoic acid, a citric acid, a maleic acid, a malonic acid, a mandelicacid, a cinnamic acid, a stearic acid, a palmitic acid, a glycolic acid,a glutamic acid, a tartaric acid, a gluconic acid, a lactic acid, afumaric acid, a lactobionic acid, an ascorbic acid, a salicylic acid, oran acetylsalicylic acid (aspirin), a salt with an amino acid such asglycine, alanine, vanillin, isoleucin, serine, cysteine, cystine, anasparaginic acid, glutamine, lysine, arginine, tyrosine, or proline, asalt with a sulfonic acid such as a methanesulfonic acid, anethanesulfonic acid, a benzenesulfonic acid, or a toluenesulfonic acid,a metal salt by a reaction with an alkali metal such as sodium orpotassium, a salt with an ammonium ion, or the like.

The arylethene derivative of the present invention may exist in asolvated form, for example, a hydrated form and a non-solvated form, andthe solvate of the arylethene derivative according to the presentinvention includes all solvated forms having a pharmaceutical activity.That is, the arylethene derivative of the present invention is dissolvedin water-compatible solvent such as methanol, ethanol, acetone, and1,4-dioxane, and then a free acid or a free base is added thereto toperform crystallization or recrystallization, thereby forming a solvateincluding a hydrate. Accordingly, as a novel compound of the presentinvention, stoichiometric solvates including hydrates may be included,in addition to a compound containing various amounts of water which maybe prepared by a method such as lyophilization.

The arylethene derivative of the present invention may have a chiralcenter, and exist as a racemate, a racemic mixture, and individualenantiomer or diastereomer. These isomers may be separated or resolvedby a common method, and an optional predetermined isomer may be obtainedby a common synthesis method or stereospecific or asymmetric synthesis.These isomer forms and mixtures thereof are all included in the scope ofthe present invention.

The arylethene derivative of the present invention may be administeredin the form of a prodrug which is decomposed in a human or animal bodyto provide the compound of the present invention. The prodrug may beused for modifying or improving a physical and (or) pharmacokineticprofile of a parent compound, and may be formed when the parent compoundcontains an appropriate group or substituent which may be derived toform the prodrug.

In addition, the pharmaceutical composition of the present invention maybe formulated into a conventional preparation in the pharmaceuticalfield, for example, a preparation for oral administration such as atablet, a pill, a hard/soft capsule, a liquid, a suspension, anemulsion, syrup, granules, and elixirs, or a preparation for parenteraladministration of a sterile aqueous or oily solvent for intravenous,subcutaneous, sublingual, intramuscular, or intradermal administration,by adding conventional non-toxic pharmaceutically acceptable carrier,excipient, and the like to the arylethene derivative represented byChemical Formula 1, or the prodrug, solvate, stereoisomer, orpharmaceutically acceptable salt thereof.

The pharmaceutically acceptable carrier which may be used in thepharmaceutical composition of the present invention is commonly used informulation, and includes lactose, dextrose, sucrose, sorbitol,mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin,calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone,cellulose, water, syrup, methyl cellulose, hydroxybenzoate,propylhydroxybenzoate, talc, magnesium stearate, and/or mineral oil, andthe like, but not limited thereto.

The excipient which may be used in the pharmaceutical composition of thepresent invention may be a sweetener, a binder, a solubilizer, asolubilizing aid, a wetting agent, an emulsifier, an isotonic agent, anadsorbent, a disintegrant, an antioxidant, a preservative, a lubricant,a filler, a fragrance, or the like, and a ratio and properties of theexcipient may be determined by solubility and chemical properties of aselected tablet, a selected administration route, and standardpharmaceutical practice. An example of the excipient may includelactose, dextrose, sucrose, mannitol, sorbitol, cellulose, glycine,silica, talc, stearic acid, sterin, magnesium stearate, magnesiumaluminum silicate, starch, gelatin, tragacanth gum, alginic acid, sodiumalginate, methyl cellulose, sodium carboxymethyl cellulose, agar, water,ethanol, polyethyleneglycol, polyvinylpyrrolidone, sodium chloride,calcium chloride, orange essence, strawberry essence, vanilla flavor, orthe like.

In addition, the pharmaceutical composition of the present invention maybe formulated into a parenteral administration form, and in this case,intravenous administration, intraperitoneal administration,intramuscular administration, subcutaneous administration, topicaladministration, or the like may be used, and ocular administration orthe like may be used, in that the composition is a therapeutic agent forretinopathy, but not limited thereto. Here, in order to be formulatedinto a formulation for parenteral administration, the pharmaceuticalcomposition may be produced into a solution or suspension by mixing theactive ingredient, that is, the arylethene derivative of ChemicalFormula 1, or the prodrug, solvate, stereoisomer, or pharmaceuticallyacceptable salt thereof with water together with a stabilizer or abuffer, and the solution or suspension may be produced into a unitdosage form of an ampoule or vial.

In addition, the pharmaceutical composition of the present invention maybe sterilized, or further include an adjuvant such as a preservative, astabilizer, a hydrating agent or an emulsifying accelerator, a salt forregulating osmotic pressure, and/or a buffer, and other therapeuticallyuseful materials, and may be formulated according to a conventionalmethod of mixing, granulating or coating.

In addition, a dosage of the arylethene derivative represented byChemical Formula 1, or the prodrug, solvate, stereoisomer, orpharmaceutically acceptable salt thereof as the active ingredient in thepharmaceutical composition according to the present invention formammals including a human may be varied depending on the age, weight,gender, dosage form, health status, and disease severity of a patient.Generally, an effective amount of 0.001 to 100 mg/kg (body weight),preferably 0.01 to 100 mg/kg (body weight) per day may be included inthe pharmaceutical composition, and the pharmaceutical composition maybe divided into once or twice per day, and administered via an oral orparenteral route. However, the amount may be increased or decreaseddepending on the administration route, severity of the disease, gender,weight, age, and the like, and thus, the administration amount in no waylimits the scope of the present invention.

In addition, the present invention provides a pharmaceutical compositionfor treating thyroid cancer comprising the arylethene derivative ofChemical Formula 1 which may specifically and significantly inhibit anERRγ transcriptional activity, or the prodrug, solvate, stereoisomer, orpharmaceutically acceptable salt thereof, and a pharmaceuticallyacceptable carrier, and used in combination of radioactive iodine.

In addition, the present invention provides a kit for treating thyroidcancer comprising the arylethene derivative of Chemical Formula 1 whichmay specifically and significantly inhibit an ERRγ transcriptionalactivity, or the prodrug, solvate, stereoisomer, or pharmaceuticallyacceptable salt thereof, and radioactive iodine.

The arylethene derivative according to the present invention regulatesexpression of endogenous ERRγ protein to regulate mitogen-activatedprotein (MAP) kinase, and improves a sodium iodide symporter (NIS)function to increase membrane-localized NIS, thereby increasing aradioactive iodine uptake when treating thyroid cancer.

Hereinafter, the present invention will be described in more detail byway of the Examples and the Experimental Examples. However, thefollowing Examples and Experimental

Examples are only illustrative of the present invention, and do notlimit the disclosure of the present invention in any way.

[Example 1] Preparation of (E)-tert-butyl4-(2-(4-(5-methoxy-5-oxo-2-phenyl-1-(4-(pivaloyloxy)phenyl)pent-1-en-1-yl)phenoxy)ethyl)piperazine-1-carboxylate(6a)

Step 1: Preparation of[4-[4-(2,2-dimethylpropanoyloxy)benzoyl]phenyl]2,2-dimethylpropanoate(A-1)

4,4-Hydroxybenzophenone (10 g, 46.6 mmol) was dissolved in 140 mL ofdichloromethane and 40 mL of tetrahydrofuran, pivaloyl chloride (19.7 g,186 mmol) and triethylamine (26 mL, 186 mmol) were slowly added thereto,and then a reaction was carried out at room temperature for 12 hours.Saturated sodium hydrogen carbonate and dichloromethane were furtheradded to the reaction solution and an organic layer was extracted. Theorganic layer was dried with anhydrous Na₂SO₄ and filtered. The solventwas distilled under reduced pressure to obtain a residue, which waspurified using column chromatography, thereby obtaining 16 g of thedesired compound A-1 (91%).

Step 2: Preparation of[4-(4-hydroxybenzoyl)phenyl]2,2-dimethylpropanoate (A-2)

Compound A-1 (12.4 g, 32.3 mmol) and potassium carbonate (2.2 g, 16.2mmol) were dissolved in methanol (360 mL) and dichloromethane (60 mL),and a reaction was carried out at room temperature for 12 hours. A 1 Maqueous citric acid solution (16.2 mL, 16.2 mmol) was added to thereaction solution, and extraction was performed with ethyl acetate. Theorganic layer was dried with anhydrous Na₂SO₄ and filtered. The solventwas distilled under reduced pressure to obtain a residue, which waspurified using column chromatography, thereby obtaining 5.6 g of thedesired compound A-2 (58%).

Step 3: Preparation of(E)-5-[4-(2,2-dimethylpropanoyloxy)phenyl]-5-(4-hydroxyphenyl)-4-phenyl-pent-4-enoate(A-3)

Zinc (8.8 g, 134 mmol) was added to tetrahydrofuran (130 mL), thetemperature was lowered to 0° C., and titanium chloride (7.35 mL, 67mmol) was slowly added thereto. The reaction solution was heated at 60°C. for 2 hours, and then compound A-2 (5 g, 16.8 mmol) andmethyl-3-benzoylpropionate (4.8 g, 25.1 mmol) were added thereto. Thereaction solution was heated at 50° C. for 1 hour. The reaction mixturewas poured into a 10% aqueous potassium carbonate solution, stirring wasperformed for 30 minutes, and filtration was performed using celite. Thefiltrate was extracted with ethyl acetate and the organic layer wasdried with anhydrous Na₂SO₄ and filtered. The solvent was distilledunder reduced pressure to obtain a residue, which was purified usingcolumn chromatography, thereby obtaining 5.4 g of the desired compoundA-3 (70%).

Step 4: Preparation of (E)-tert-butyl4-(2-(4-(5-methoxy-5-oxo-2-phenyl-1-(4-(pivaloyloxy)phenyl)pent-1-en-1-yl)phenoxy)ethyl)piperazine-1-carboxylate(A-4)

To dichloromethane (3 mL), compound A-3 (0.05 g, 0.11 mmol),2-(4-(tert-butyloxycarbonyl)piperazin-1-yl)ethanol (30 mg, 0.13 mmol),and triphenylphosphine (86 mg, 0.33 mmol) were added, the temperaturewas lowered to 0° C., and diisopropyl azodicarboxylate (0.064 mL, 0.33mmol) was slowly added thereto. After 15 minutes, the temperature wasraised to room temperature, and stirring was performed for 12 hours.Water and ethyl acetate were further added to the reaction solution andan organic layer was extracted. The organic layer was dried withanhydrous Na₂SO₄ and filtered. The solvent was distilled under reducedpressure to obtain a residue, which was purified using columnchromatography, thereby obtaining 73 mg of the desired compound A-4(99%).

Step 5: Preparation of(Z)-4-(5-hydroxy-1-(4-(2-(4-(tert-Butyloxycarbonyl)piperazin-1-yl)ethoxy)phenyl)-2-phenylpent-1-en-1-yl)phenol(6a)

Compound C (0.34 g, 0.05 mmol) was added to tetrahydrofuran (10 mL), thetemperature was lowered to 0° C., and 1 M lithium aluminum hydride(LiAlH₄, 1.5 mL, 1.51 mmol) was slowly added thereto. The temperaturewas raised to room temperature, and stirring was performed for 1 hour.Water and ethyl acetate were further added to the reaction solution andan organic layer was extracted. The organic layer was dried withanhydrous Na₂SO₄ and filtered. The solvent was distilled under reducedpressure to obtain a residue, which was purified using columnchromatography, thereby obtaining 0.28 g of the desired compound 6a(99%).

Examples 2 to 13

Compounds 6b to 6m were prepared according to the process of Example 1.Compounds 6b to 6m were prepared by the same process, except that instep 4 of Example 1, 2-(4-(tert-butyloxycarbonyl)piperazin-1-yl)ethanolwas replaced with different ethanol. Identification data of thethus-prepared compounds 6a to 6m is shown in the following Table 1.

TABLE 1

Cmpd Example No. R Identification data 1 6a

¹H-NMR (CD₃OD, 400 MHz) δ 7.22-7.11 (m, 7H), 7.05 (d, J = 4.1 Hz, 2H),6.80 (d, J = 6.5 Hz, 2H), 3.77 (s, 4H), 3.43 (m, 6H), 2.56 (m, 2H), 1.59(m, 2H), 1.49 (s, 9H). MS (ESI) m/z: 515 [M + H]⁺. 2 6b

¹H-NMR (CD₃OD, 400 MHz) δ 7.19-7.08 (m, 5H), 7.05-7.02 (m, 2H),6.85-6.78 (m, 4H), 6.66-6.63 (m, 2H), 4.19 (t, J = 4.6 Hz, 2H),3.50-3.37 (m, 12H), 2.94 (s, 3H), 2.53 (t, J = 2.8 Hz, 2H), 1.53 (m,2H). MS (ESI) m/z: 473 [M + H]⁺. 3 6c

¹H-NMR (DMSO-d₆, 400 MHz) δ 7.17 (m, 2H), 7.10 (m, 3H), 6.97 (d, J = 8.5Hz, 2H), 6.75 (m, 4H), 6.66 (d, J = 8.8 Hz, 2H), 4.24 (t, J = 4.3 Hz,2H), 3.93 (m, 2H), 3.74 (m, 2H), 3.24 (t, J = 6.7 Hz, 2H), 3.15 (s, 2H),2.54 (s, 4H), 2.39 (m, 2H), 1.38 (m, 2H). MS (ESI) m/z: 460 [M + H]⁺. 46d

¹H-NMR (CD₃OD, 400 MHz) δ 7.18-7.09 (m, 5H), 7.04 (d, J = 8.6 Hz, 2H),6.85 (d, J = 8.8 Hz, 2H), 6.79 (d, J = 8.5 Hz, 2H), 6.68 (d, J = 8.8 Hz,2H), 4.25 (t, J = 4.8 Hz, 2H), 3.58 (m, 2H), 3.50 (t, J = 4.9 Hz, 2H),3.45 (t, J = 6.8 Hz, 2H), 3.04 (m, 2H), 2.54 (m, 2H), 1.96 (m, 2H), 1.82(m, 3H), 1.55 (m, 3H). MS (ESI) m/z: 458 [M + H]⁺. 5 6e

¹H-NMR (CD₃OD, 400 MHz) δ 7.18-7.09 (m, 5H), 7.04 (d, J = 8.5 Hz, 2H),6.85 (d, J = 8.7 Hz, 2H), 6.78 (d, J = 8.5 Hz, 2H), 6.68 (d, J = 8.8 Hz,2H), 4.21 (t, J = 4.8 Hz, 2H), 3.69 (m, 2H), 3.59 (t, J = 4.9 Hz, 2H),3.43 (t, J = 6.8 Hz, 2H), 3.18 (m, 2H), 2.54 (m, 2H), 2.18 (m, 2H), 2.04(m, 2H), 1.56 (m, 2H). MS (ESI) m/z: 444 [M + H]⁺. 6 6f

¹H-NMR (CD₃OD, 400 MHz) δ 7.17-1.08 (m, 5H), 7.04 (d, J = 8.5 Hz, 2H),6.84 (d, J = 8.6 Hz, 2H), 6.78 (d, J = 8.5 Hz, 2H), 6.68 (d, J = 8.7 Hz,2H), 4.20 (t, J = 4.5 Hz, 2H), 3.91 (t, J = 5.5 Hz, 2H), 3.50 (m, 4H),3.44 (t, J = 6.8 Hz, 2H), 2.53 (m, 2H), 1.55 (m, 2H). MS (ESI) m/z: 416[M + H]⁺. 7 6g

¹H-NMR (CD₃OD, 400 MHz) δ 7.18-7.09 (m, 5H), 7.04 (d, J = 8.5 Hz, 2H),6.80 (m, 4H), 6.62 (d, J = 8.6 Hz, 2H), 4.09-3.97 (m, 2H), 3.67 (m, 1H),3.52 (m, 1H), 3.43 (t, J = 6.7 Hz, 2H), 3.16 (m, 1H), 2.94 (s, 3H), 2.53(t, J = 7.8 Hz, 2H), 2.38 (m, 2H), 2.19- 2.01 (m, 3H), 1.85 (m, 1H),1.54 (m, 2H). MS (ESI) m/z: 458 [M + H]⁺. 8 6h

¹H-NMR (CD₃OD, 400 MHz) δ 7.17-7.08 (m, 5H), 7.05 (d, J = 8.4 Hz, 2H),6.85 (d, J = 8.7 Hz, 2H), 6.79 (d, J = 8.4 Hz, 2H), 6.69 (d, J = 8.7 Hz,2H), 4.30 (m, 1H), 4.09 (m, 1H), 3.82 (m, 1H), 3.68 (m, 1H), 3.43 (t, J= 6.7 Hz, 2H), 3.21 (m, 1H), 3.02 (s, 3H), 2.54 (m, 2H), 2.35 (m, 1H),2.20 (m, 1H), 2.02 (m, 2H), 1.56 (m, 2H). MS (ESI) m/z: 444 [M + H]⁺. 96i

¹H-NMR (CD₃OD, 400 MHz) δ 7.17-7.08 (m, 5H), 7.05 (d, J = 8.5 Hz, 2H),6.85 (d, J = 8.8 Hz, 2H), 6.79 (d, J = 8.5 Hz, 2H), 6.69 (d, J = 8.8 Hz,2H), 4.28 (m,. 1H), 4.09 (m, 1H), 3.83 (m, 1H), 3.69 (m, 1H), 3.43 (t, J= 6.7 Hz, 2H), 3.21 (m, 1H), 3.02 (s, 3H), 2.54 (m, 2H), 2.35 (m, 1H),2.22 (m, 1H), 1.99 (m, 2H), 1.56 (m, 2H). MS (ESI) m/z: 444 [M + H]⁺. 106j

¹H-NMR (CD₃OD, 400 MHz) δ 7.16-7.07 (m, 5H), 7.02 (d, J = 8.6 Hz, 2H),6.78 (m, 4H), 6.60 (d, J = 8.8 Hz, 2H), 4.03 (m, 2H), 3.67 (m, 1H), 3.51(m, 1H), 3.41 (t, J = 6.8 Hz, 2H), 3.15 (m, 1H), 2.92 (s, 3H), 2.51 (m,2H), 2.38 (m, 1H), 2.08 (m, 4H), 1.84 (m, 1H), 1.55 (m, 2H). MS (ESI)m/z: 458 [M + H]⁺. 11 6k

¹H-NMR (CD₃OD, 400 MHz) δ 7.16-7.07 (m, 5H), 7.02 (d, J = 6.6 Hz, 2H),6.78 (m, 4H), 6.60 (d, J = 8.8 Hz, 2H), 4.01 (m, 2H), 3.67 (m, 1H), 3.50(m, 1H), 3.41 (t, J = 6.7 Hz, 2H), 3.15 (m, 1H), 2.92 (s, 3H), 2.51 (m,2H), 2.37 (m, 1H), 2.07 (m, 4H), 1.84 (m, 1H), 1.54 (m, 2H). MS (ESI)m/z: 458 [M + H]⁺. 12 6l

¹H-NMR (CD₃OD, 400 MHz) δ 7.18-7.09(m, 5H), 7.05 (d, J = 8.6 Hz, 2H),6.85 (d, J = 8.8 Hz, 2H), 6.79 (d, J = 8.6 Hz, 2H), 6.71 (d, J = 8.8 Hz,2H), 3.63 (m, 1H), 3.49 (m, 2H), 3.43 (t, J = 6.8 Hz, 2H), 3.25 (m, 1H),3.04 (m, 1H), 2.87 (s, 3H), 2.54 (m, 2H), 2.04 (m, 2H), 2.04 (m, 2H),1.79 (m, 1H), 1.66 (m, 1H), 1.55 (m, 2H). MS (ESI) m/z: 444 [M + H]⁺. 136m

¹H-NMR (CD₃OD, 400 MHz) δ 7.17-7.09 (m, 5H), 7.04 (d, J = 2H), 6.84 (m,2H), 6.78 (m, 2H), 6.71 (m, 2H), 3.62 (m, 1H), 3.43 (m, 4H), 3.26 (m,1H), 3.04 (m, 1H), 2.87 (s, 3H), 2.53 (m, 2H), 2.05 (m, 2H), 1.77 (m,1H), 1.69 (m, 1H), 1.54 (m, 2H). MS (ESI) m/z: 444 [M + H]⁺.

[Example 14] Preparation of(Z)-4-(5-hydroxy-2-phenyl-1-(4-(2-(piperazin-1-yl)ethoxy)phenyl)pent-1-en-1-yl)phenol2hydrochloride salt (7a)

Compound 6a (28 mg, 0.05 mmol) was added to dichloromethane (5 mL), thetemperature was lowered to 0° C., and trifluoroacetic acid (0.08 mL,1.00 mmol) was added thereto. The temperature was raised to roomtemperature, and stirring was performed for 1 hour. Water and ethylacetate were further added to the reaction solution and an organic layerwas extracted. The organic layer was dried with anhydrous Na₂SO₄ andfiltered. The solvent was distilled under reduced pressure to obtain aresidue, which was purified using column chromatography and thendissolved in methanol:dichloromethane (1:1), the temperature was loweredto 0° C., a 1M aqueous HCl solution was slowly added thereto, anddistillation under reduced pressure was performed, thereby obtaining 4mg of the desired compound 7a (17%).

¹H-NMR (CD₃OD, 400 MHz) δ 7.16-7.07 (m, 5H), 7.01 (d, J=8.6 Hz, 2H),6.83 (d, J=8.8 Hz, 2H), 6.76 (d, J=8.6 Hz, 2H), 4.31 (t, J=4.2 Hz, 2H),3.66 (m, 10H), 3.41 (t, J=6.8 Hz, 2H), 2.51 (m, 2H), 1.54 (m, 2H). MS(ESI) m/z: 459 [M+H]⁺.

[Example 15] Preparation of(E)-5-(4-(2-(aziridin-1-yl)ethoxy)phenyl)-5-(4-bromophenyl)-4-phenylpent-4-en-1-olhydrochloride salt (13a)

Step 1: Preparation of (4-bromophenyl)(4-methoxyphenyl)methanone (B-1)

4-Bromobenzoyl chloride (8.2 g, 50.9 mmol) and aluminum chloride (6.1 g,50.9 mmol) were dissolved in dichloromethane (90 mL), and anisole (5 g,46.2 mmol) was slowly added thereto. Stirring was performed for 3 hours,the temperature was lowered to 0° C., and 1N HCl (50 mL) was addedthereto. Ethyl acetate was added to extract an aqueous layer, which wasdried with anhydrous Na₂SO₄ and filtered. The solvent was distilledunder reduced pressure to obtain a residue, which was purified usingcolumn chromatography, thereby obtaining 11 g of the desired compoundB-1 (99%).

Step 2: Preparation of (4-bromophenyl)(4-hydroxyphenyl)methanone (B-2)

Compound B-1 (10 g, 34.3 mmol) was added to toluene (80 mL), thetemperature was lowered to 0° C., and aluminum chloride (11.5 g, 86mmol) was slowly added thereto. Heating was performed at 70° C. for 4hours. The reaction solution was cooled to room temperature, 1 Nhydrochloric acid was added thereto, ethyl acetate was added thereto,and extraction was performed. The organic layer was dried with anhydrousNa₂SO₄ and filtered. The solvent was distilled under reduced pressure toobtain a residue, which was purified using column chromatography,thereby obtaining 8.1 g of the desired compound B-2 (85%).

Step 3: Preparation of (E)-methyl5-(4-bromophenyl)-5-(4-hydroxyphenyl)-4-phenylpent-4-enoate (B-3)

0.61 g of the desired compound B-3 (39%) was obtained by the sameprocess as step 3 of Example 1

Step 4: Preparation of (E)-methyl5-(4-(2-(aziridin-1-yl)ethoxy)phenyl)-5-(4-bromophenyl)-4-phenylpent-4-enoate(B-4)

44 mg of the desired compound B-4 (54%) was obtained, using compound B-3and 2-(aziridin-1-yl)ethanol by the same process as step 4 of Example 1.

Step 5: Preparation of(E)-5-(4-(2-(aziridin-1-yl)ethoxy)phenyl)-5-(4-bromophenyl)-4-phenylpent-4-en-1-olhydrochloride salt (13a)

Compound B-4 (44 mg, 0.09 mmol) was added to tetrahydrofuran (2 mL), thetemperature was lowered to 0° C., and 1 M diisobutylaluminum hydride(0.26 mL, 0.26 mmol) was slowly added thereto. The temperature wasraised to room temperature, and stirring was performed for 1 hour. Waterand ethyl acetate were further added to the reaction solution and anorganic layer was extracted. The organic layer was dried with anhydrousNa₂SO₄ and filtered. The solvent was distilled under reduced pressure toobtain a residue, which was purified using column chromatography,thereby obtaining 0.3 mg of the desired compound 13a (0.7%).

Examples 16 to 18

Compounds 13b to 13d were prepared according to the process of Example15. Compounds 13b to 13d were prepared by the same process, except thatin step 4 of Example 15, 2-(aziridin-1-yl)ethanol was replaced withdifferent ethanol. Identification data of the thus-prepared compounds13a to 13d is shown in the following Table 2.

TABLE 2

Cmpd Example No. R Identification data 15 13a

¹H-NMR(CD₃OD, 400 MHz) δ 7.51 (d, J = 8.4 Hz, 2H), 7.16-7.09 (m, 7H),6.83 (d, J = 8.8 Hz, 2H), 6.69 (d, J = 8.8 Hz, 2H), 4.18 (t, J = 4.7 Hz,2H), 3.89 (t, J = 5.6 Hz, 2H), 3.49 (m, 4H), 3.41 (t, J = 6.6 Hz, 2H),2.49 (m, 2H), 1.53 (m, 2H). MS (ESI) m/z: 479 [M + H]⁺. 16 13b

¹H-NMR(CD₃OD, 400 MHz) δ 7.51 (d, J = 8.4 Hz, 2H), 7.16-7.09 (m, 7H),6.84 (d, J = 8.8 Hz, 2H), 6.70 (d, J = 8.8 Hz, 2H), 4.27 (m, 1H), 4.08(m, 1H), 3.80 (m, 1H), 3.66 (m, 1H), 3.41 (t, J = 6.5 Hz, 2H), 3.19 (m,1H), 3.00 (s, 3H), 2.50 (m, 2H), 2.33 (m, 1H), 2.20 (m, 1H), 1.98 (m,2H), 1.52 (m, 2H). MS (ESI) m/z: 507 [M + H]⁺. 17 13c

¹H-NMR(CD₃OD, 400 MHz) δ 7.52 (d, J = 8.4 Hz, 2H), 7.17-7.12 (m, 7H),6.84 (d, J = 8.7 Hz, 2H), 6.71 (d, J = 8.6 Hz, 2H), 3.61 (m, 1H), 3.41(m, 4H), 3.23 (m, 1H), 3.02 (m, 1H), 2.85 (s, 3H), 2.49 (m, 2H), 2.02(m, 2H), 1.77 (m, 1H), 1.65 (m, 2H), 1.53 (m, 2H). MS (ESI) m/z: 507[M + H]⁺. 18 13d

¹H-NMR(CD₃OD, 400 MHz) δ 7.52 (d, J = 8.4 Hz, 2H), 7.15 (m, 7H), 6.80(m, 2H), 6.64 (m, 2H), 4.05 (m, 1H), 3.50 (m, 2H), 3.43 (m, 2H), 3.15(m, 2H), 2.92 (m, 3H), 2.50 (m, 2H), 2.09 (m, 6H), 1.55 (m, 2H). MS(ESI) m/z: 520 [M + H]⁺.

[Example 19] Preparation of(E)-5-(4-(2-(aziridin-1-yl)ethoxy)phenyl)-5-(4-bromophenyl)-4-phenylpent-4-en-1-olhydrochloride salt (18t)

Step 1: Preparation of methyl 5-(4-(pivaloyloxy)phenyl)pent-4-ynoate(C-1)

4-Iodophenyl pivalate (2 g, 6.6 mmol), copper (I) chloride (0.13 g, 0.66mmol), bis(triphenylphosphine)palladium (II) dichloride (PdCl₂(PPh₃)₂,0.23 g, 0.33 mmol), and methyl pent-4-ynoate (0.74 g, 0.66 mmol) weredissolved in trimethylamine (15 mL), and the reaction was carried out at50° C. for 12 hours. The reaction solution was concentrated underreduced pressure, and 1.1 g of the desired compound C-1 (58%) wasobtained using column chromatography.

Step 2: Preparation of (E)-tert-butyl3-(4-(5-methoxy-5-oxo-2-phenyl-1-(4-(pivaloyloxy)phenyl)pent-1-en-1-yl)phenyl)azetidin-1-carboxylate(C-2)

tert-Butyl3-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaboran-2-yl)phenyl)azetidin-1-carboxylate(0.27 g, 0.75 mmol), compound C-1 (0.14 g, 0.5 mmol), and iodobenzene(84 μL, 0.75 mmol) were dissolved in DMF (8 mL) and water (4 mL), 0.025M PdCl₂(PhCN)₂ (0.2 mL, 5 μmol) was added thereto, and heating wasperformed at 45° C. for 10 minutes. Cesium carbonate (0.24 g, 0.75 mmol)was added thereto, and heating was performed at 45° C. for 12 hours.When the reaction was completed, brine and ethyl acetate was furtheradded to the reaction solution, and an organic layer was extracted. Theorganic layer was dried with anhydrous Na₂SO₄ and filtered. The solventwas distilled under reduced pressure to obtain a residue, which waspurified using column chromatography, thereby obtaining 81 mg of thedesired compound C-2 (27%).

Step 3: Preparation of tert-butyl(E)-3-(4-(5-hydroxy-1-(4-hydroxyphenyl)-2-phenylpent-1-en-1-yl)phenyl)azetidine-1-carboxylate(18t)

Compound C-2 (0.021 mmol) was added to tetrahydrofuran (2 mL), thetemperature was lowered to 0° C., and 1 M lithium aluminum hydride,diisobutylaluminum hydride, or lithium borohydride (0.024 mL, 0.024mmol) was added thereto. The temperature was raised to room temperature,and stirring was performed for 1 hour. Water and ethyl acetate werefurther added to the reaction solution and an organic layer wasextracted. The organic layer was dried with anhydrous Na₂SO₄ andfiltered. The solvent was distilled under reduced pressure to obtain aresidue, which was purified using column chromatography and thendissolved in methanol:dichloromethane (1:1), the temperature was loweredto 0° C., a 1M aqueous HCl solution was slowly added thereto, anddistillation under reduced pressure was performed, thereby obtaining 24mg of the desired compound 18t (78%).

Examples 20 to 39

Compounds 18a to 18s and 18u were prepared using the process of Example19. Identification data of the thus-prepared compounds 18a to 18u isshown in the following Table 3.

TABLE 3

          Example         Cmpd No.

          R           Identification data 19 18t

¹H-NMR (CD₃OD, 400 MHz) δ 7.18-7.10 (m, 5H), 7.05 (d, J = 8.4 Hz, 2H),6.98 (d, J = 8.2 Hz, 2H), 6.88 (d, J = 8.2 Hz, 2H), 6.79 (d, J = 8.4 Hz,2H), 4.25 (t, J = 8.4 Hz, 2H), 3.81 (t, J = 6.6 Hz, 2H), 3.66 (m, 1H),3.43 (t, J = 6.8 Hz, 2H), 2.55 (m, 2H), 1.57 (m, 2H), 1.45 (s, 9H). MS(ESI) m/z: 386 [M + H]⁺. 20 18a

¹H-NMR(CD₃OD, 400 MHz) δ 7.14-7.07 (m, 5H), 7.02 (d, J = 8.0 Hz, 2H),6.78 (m, 4H), 6.69 (d, J = 8.3 Hz, 2H), 3.76 (m, 2H), 3.52 (m, 3H), 3.41(t, J = 6.4 Hz, 2H), 3.23 (m, 2H), 2.96 (m, 2H), 2.51 (m, 2H), 1.53 (m,2H), 1.39 (d, J = 6.5 Hz, 6H). MS (ESI) m/z: 457 [M + H]⁺. 21 18b

¹H-NMR(CD₃OD, 400 MHz) δ 7.18-7.13 (m, 5H), 7.08 (m, 4H), 6.79 (d, J =8.7 Hz, 2H), 6.67 (d, J = 8.7 Hz, 2H), 3.75 (m, 2H), 3.53 (m, 3H), 3.39(t, J = 6.8 Hz, 2H), 3.21 (m, 2H), 2.92 (m, 2H), 2.48 (m, 2H), 2.35 (s,3H), 1.53 (m, 2H), 1.38 (d, J = 6.6 Hz, 6H). MS (ESI) m/z: 455 [M + H]⁺.22 18c

¹H-NMR(CD₃OD, 400 MHz) δ 8.25 (d, J = 8.7 Hz, 2H), 7.47 (d, J = 8.7 Hz,2H), 7.21-7.13 (m, 5H), 6.81 (d, J = 8.7 Hz, 2H), 6.71 (d, J = 8.8 Hz,2H), 3.74 (m, 2H), 3.54 (m, 2H), 3.42 (t, J = 6.4 Hz, 2H), 3.19 (t, J =9.8 Hz, 2H), 2.93 (m, 5H), 2.50 (m, 2H), 1.55 (m, 2H). MS (ESI) m/z: 458[M + H]⁺. 23 18d

¹H-NMR (CD₃OD, 400 MHz) δ 7.20-7.07 (m, 7H), 7.04 (d, J = 8.5 Hz, 2H),6.83 (d, J = 8.7 Hz, 2H), 6.78 (d, J = 8.5 Hz, 2H), 6.73 (d, J = 8.7 Hz,2H), 6.66 (d, J = 8.6 Hz, 2H), 3.74 (m, 2H), 3.57 (m, 2H), 3.42 (t, J =6.7 Hz, 2H), 3.33 (m, 2H), 3.23 (m, 2H), 2.95 (s, 3H), 2.53 (m, 2H),1.55 (m, 2H). MS (ESI) m/z: 429 [M + H]⁺. 24 18e

¹H-NMR(CD₃OD, 400 MHz) δ 7.49 (d, J = 8.4 Hz, 2H), 7.14-7.12 (m, 7H),6.72 (d, J = 8.8 Hz, 2H), 6.61 (d, J = 8.9 Hz, 2H), 3.40 (t, J = 6.6 Hz,2H), 3.07 (m, 4H), 2.55 (m, 4H), 2.47 (m, 2H), 2.31 (s, 3H), 1.52 (m,2H). MS (ESI) m/z: 492 [M + H]⁺. 25 18f

¹H-NMR(CD₃OD, 400 MHz) δ 7.19-7.09 (m, 9H), 6.81 (d, J = 8.6 Hz, 2H),6.68 (d, J = 8.8 Hz, 2H), 3.70 (m, 2H), 3.55 (m, 2H), 3.42 (t, J= 6.8Hz, 2H), 3.20 (m, 2H), 2.00 (m, 2H), 2.93 (s, 3H), 2.50 (m, 2H), 2.36(s, 3H), 1.55 (m, 2H). MS (ESI) m/z: 427 [M + H]⁺. 26 18g

¹H-NMR(CD₃OD, 400 MHz) δ 8.25 (d, J = 8.7 Hz, 2H), 7.47 (d, J = 8.7 Hz,2H), 7.21-7.13 (m, 5H), 6.81 (d, J = 8.7 Hz, 2H), 6.72 (d, J = 8.8 Hz,2H), 3.78 (m, 2H), 3.53 (m, 3H), 3.42 (t, J = 6.5 Hz, 2H), 3.21 (m, 2H),2.93 (m, 2H), 2.50 (m, 2H), 1.55 (m, 2H), 1.39 (d, J = 6.6 Hz, 6H). MS(ESI) m/z: 486 [M + H]⁺. 27 18h

¹H-NMR(CD₃OD, 400 MHz) δ 7.49 (d, J = 8.4 Hz, 2H), 7.16-7.11 (m, 7H),6.74 (d, J = 8.8 Hz, 2H), 6.63 (d, J = 8.8 Hz, 2H), 3.40 (t, J = 6.6 Hz,2H), 3.16 (m, 4H), 2.96 (m, 5H), 2.47 (m, 2H), 1.54 (m, 2H), 1.19 (d, J= 6.6 Hz, 6H). MS (ESI) m/z: 520 [M + H]⁺. 28 18i

¹H-NMR(CD₃OD, 400 MHz) δ 7.16-7.07 (m, 5H), 7.01 (d, J = 7.6 Hz, 2H),6.77 (m, 4H), 6.67 (d, J = 8.5 Hz, 2H), 3.73 (m, 2H), 3.58 (m, 2H), 3.41(t, J = 6.8 Hz, 2H), 3.23 (m, 2H), 3.14 (m, 2H), 2.93 (m, 2H), 2.51 (m,2H), 1.53 (m, 2H), 1.37 (t, J = 7.3 Hz, 3H). MS (ESI) m/z: 443 [M + H]⁺.29 18j

¹H-NMR(CD₃OD, 400 MHz) δ 7.19-7.15 (m, 5H), 7.10 (m, 4H), 6.8 (d, J =8.8 Hz, 2H), 6.68 (d, J = 8.9 Hz, 2H), 3.75 (m, 2H), 3.60 (m, 2H), 3.41(t, J = 6.8 Hz, 2H), 3.25 (d, J = 7.4 Hz, 2H), 3.14 (m, 2H), 2.93 (m,2H), 2.50 (m, 2H), 2.37 (s, 3H), 1.55 (m, 2H), 1.38 (t, J = 7.3 Hz, 2H).MS (ESI) m/z: 441 [M + H]⁺. 30 18k

¹H-NMR(CD₃OD, 400 MHz) δ 7.19-7.09 (m, 6H), 6.81 (d, J = 8.7 Hz, 2H),6.69 (m, 4H), 6.63 (m, 1H), 3.76 (m, 2H), 3.53 (m, 3H), 3.40 (t, J = 5.6Hz, 2H), 3.21 (m, 2H), 2.91 (m, 2H), 2.50 (m, 2H), 1.53 (m, 2H), 1.38(d, J = 6.6 Hz, 6H). MS (ESI) m/z: 457 [M + H]⁺. 31 18l

¹H-NMR(CD₃OD, 400 MHz) δ 7.17-7.07 (m, 7H), 6.89 (d, J = 8.7 Hz, 2H),6.85 (m, 2H), 6.65 (d, J = 8.8 Hz, 2H), 3.73 (m, 2H), 3.50 (m, 2H), 3.35(m, 2H), 3.20 (m, 2H), 2.91 (m, 2H), 2.39 (t, J = 7.9 Hz, 2H), 1.57 (m,2H), 1.38 (d, J = 6.6 Hz, 6H). MS (ESI) m/z: 457 [M + H]⁺. 32 18m

¹H-NMR(CD₃OD, 400 MHz) δ 7.22-7.11 (m, 7H), 7.05 (m, 4H), 6.80 (d, J =8.6 Hz, 2H), 3.77 (m, 4H), 3.42 (m, 6H), 2.55 (m, 2H), 1.59 (m, 2H),1.49 (s, 9H). MS (ESI) m/z: 515 [M + H]⁺. 33 18n

¹H-NMR(CD₃OD, 400 MHz) δ 7.49 (d, J = 8.4 Hz, 2H), 7.16-7.12 (m, 7H),6.72 (d, J = 8.8 Hz, 2H), 6.61 (d, J = 8.8 Hz, 2H), 3.47 (s, 4H), 3.39(t, J = 6.6 Hz, 2H), 2.98 (m, 4H), 2.47 (m, 2H), 1.52 (m, 2H), 1.44 (s,9H). MS (ESI) m/z: 478 [M + H]⁺. 34 18o

¹H-NMR(CD₃OD, 400 MHz) δ 7.42 (d, J = 8.7 Hz, 2H), 7.20-7.12 (m, 7H),7.07 (d, J = 8.5 Hz, 2H), 6.82 (d, J = 8.7 Hz, 2H), 4.07 (m, 4H), 3.62(m, 4H), 3.44 (t, J = 6.6 Hz, 2H), 2.56 (m, 2H), 1.57 (m, 2H). MS (ESI)m/z: 416 [M + H]⁺. 35 18p

¹H-NMR(CD₃OD, 400 MHz) δ 8.23 (d, J = 8.8 Hz, 2H), 7.46 (d, J = 8.6 Hz,2H), 7.22-7.12 (m, 5H), 6.66 (d, J = 8.8 Hz, 2H), 6.23 (d, J = 8.7 Hz,2H), 3.42 (t, J = 6.5 Hz, 2H), 3.27 (m, 2H), 3.13 (m, 2H), 2.48 (m, 2H),1.90 (m, 4H), 1.54 (m, 4H), 1.35 (m, 2H), 1.29 (s, 9H). MS (ESI) m/z:484 [M + H]⁺. 36 18q

¹H-NMR(CD₃OD, 400 MHz) δ 7.52 (d, J = 8.4 Hz, 2H), 7.18 (d, J = 8.4 Hz,2H), 7.17-7.10 (m, 5H), 7.03 (d, J = 8.1 Hz, 2H), 6.89 (d, J = 8.1 Hz,2H), 3.54 (s, 2H), 3.42 (t, J = 6.6 Hz, 2H), 2.52 (m, 6H), 1.62-1.48 (m,8H). MS (ESI) m/z: 492 [M + H]⁺. 37 18r

¹H-NMR(CD₃OD, 400 MHz) δ 7.38 (m, 2H), 7.31 (m, 1H), 7.22 (m, 2H), 7.07(d, J = 8.6 Hz, 2H), 6.83 (d, J = 8.6 Hz, 2H), 6.51 (s, 1H), 6.43 (s,1H), 4.03 (m, 3H), 3.35 (t, J = 6.9 Hz, 2H), 2.32 (m, 2H), 1.81 (m, 2H),1.59 (m, 6H), 1.44 (s, 9H). MS (ESI) m/z: 404 [M + H]⁺. 38 18s

¹H-NMR(CD₃OD, 400 MHz) δ 7.93 (s, 1H), 7.05-7.12 (m, 9H), 6.85 (d, J =10.0 Hz, 2H), 6.79 (d, J = 8.8 Hz, 2H), 6.47(s, 1H), 3.61 (m, 2H), 3.48(m, 4H), 3.25 (t, J = 7.6 Hz, 2H), 3.01 (t, J = 10.4 Hz, 2H), 2.59 (m,2H), 1.99 (m, 2H), 1.77-1.87 (m, 3H), 1.51-1.61 (m, 3H). MS (ESI) m/z:482 [M + H]⁺. 39 18u

¹H-NMR(CD₃OD, 400 MHz) δ 8.22 (s, 1H), 7.34 (d, J = 8.2 Hz, 2H), 6.99(m, 5H), 6.93 (d, J = 8.5 Hz, 2H), 6.82 (d, J = 8.2 Hz, 2H), 6.65 (d, J= 8.5 Hz, 2H), 4.78 (m, 2H), 3.66 (t, J = 5.8 Hz, 2H), 3.29 (t, J = 6.7Hz, 2H), 2.84 (s, 6H), 2.42 (m, 2H), 1.43 (m, 2H). MS (ESI) m/z: 469[M + H]⁺.

[Example 40] Preparation of(E)-5-(4-bromophenyl)-4-phenyl-5-(4-(piperazin-1-yl)phenyl)pent-4-en-1-ol(20a)

Step 1: Preparation of methyl(E)-5-(4-bromophenyl)-4-phenyl-5-(4-(piperazin-1-yl)phenyl)pent-4-enoate(D-2)

Compound D-1 (6 mg, 0.01 mmol) was added to dichloromethane (2 mL), thetemperature was lowered to 0° C., and trifluoroacetic acid (0.05 mL,0.65 mmol) was slowly added thereto. The temperature was raised to roomtemperature, and stirring was performed for 12 hours. Water and ethylacetate were further added to the reaction solution and an organic layerwas extracted. The organic layer was dried with anhydrous Na₂SO₄ andfiltered. The solvent was distilled under reduced pressure to obtain aresidue, which was purified using column chromatography, therebyobtaining 5 mg of the desired compound D-2 (99%).

Step 2: Preparation of(E)-5-(4-bromophenyl)-4-phenyl-5-(4-(piperazin-1-yl)phenyl)pent-4-en-1-ol(20a)

7 mg of the desired compound 20a (41%) was obtained by the same processas step 3 of Example 19, using compound D-2.

Examples 41 to 51

Compounds 20b to 201 were prepared, using the process of Example 40.Identification data of the thus-prepared compounds 20a to 201 is shownin the following Table 4.

TABLE 4

          Example         Cmpd No.

          R           Identification data 40 20a

¹H-NMR(CD₃OD, 400 MHz) δ 7.50 (d, J = 8.4 Hz, 2H), 7.17-7.09 (m, 7H),6.76 (m, 2H), 6.66 (d, J = 8.8 Hz, 2H), 3.39 (m, 2H), 3.22 (s, 8H), 2.48(m, 2H), 1.52 (m, 2H). MS (ESI) m/z: 478 [M + H]⁺. 41 20b

¹H-NMR(CD₃OD, 400 MHz) δ 7.16-7.06 (m, 9H), 6.71 (d, J = 8.8 Hz, 2H),6.64 (d, J = 8.8 Hz, 2H), 3.38 (t, J = 6.8 Hz, 2H), 3.19 (s, 8H), 2.47(m, 2H), 2.34 (s, 3H), 1.52 (m, 2H). MS (ESI) m/z: 413 [M + H]⁺. 42 20c

¹H-NMR(CD₃OD, 400 MHz) δ 7.20-7.11(m, 7H), 7.04 (m, 4H), 6.80 (d, J =8.5 Hz, 2H), 3.83 (m, 2H), 3.70 (m, 6H), 3.62 (t, J = 5.2 Hz, 2H), 2.51(m, 2H), 1.56 (m, 2H). MS (ESI) m/z: 415 [M + H]⁺. 43 20d

¹H-NMR(CD₃OD, 400 MHz) δ 7.17-7.07 (m, 9H), 7.03 (d, J = 8.3 Hz, 2H),6.92 (d, J = 8.4 Hz, 2H), 4.24 (m, 2H), 4.07 (m, 3H), 3.39 (t, J = 6.8Hz, 2H), 2.49 (m, 2H), 2.34 (s, 3H), 1.53 (m, 2H). MS (ESI) m/z: 384[M + H]⁺. 44 20e

¹H-NMR (CD₃OD, 400 MHz) δ 7.17-7.09 (m, 5H), 7.05 (m, 4H), 6.95 (d, J =7.4 Hz, 2H), 6.79 (d, J = 7.8 Hz, 2H), 4.29 (m, 2H), 4.12 (m, 3H), 3.44(t, J = 6.6 Hz, 2H), 2.55 (t, J = 7.6 Hz, 2H), 1.57 (m, 2H). MS (ESI)m/z: 386 [M + H]⁺. 45 20f

¹H-NMR(CD₃OD, 400 MHz) δ 7.51 (d, J = 8.4 Hz, 2H), 7.16-7.11 (m, 7H),7.04 (d, J = 8.2 Hz, 2H), 6.91 (d, J = 8.2 Hz, 2H), 4.18 (m, 2H), 4.02(m, 3H), 3.41 (t, J = 6.6 Hz, 2H), 2.50 (m, 2H), 1.53 (m, 2H). MS (ESI)m/z: 449 [M + H]⁺. 46 20g

¹H-NMR(CD₃OD, 400 MHz) δ 7.93 (s, 1H), 7.05-7.12 (m, 9H), 6.85 (d, J =10.0 Hz, 2H), 6.79 (d, J = 8.6 Hz 2H) 6.47(s, 1H) 3.61 (m, 2H), 3.48 (m,4H), 3.25 (t, J = 7.6 Hz, 2H), 3.01 (t, J = 10.4 Hz, 2H), 2.59 (m, 2H),1.99 (m, 2H), 1.77-1.87 (m, 3H), 1.51-1.61 (m, 3H). MS (ESI) m/z: 443[M + H]⁺. 47 20h

¹H-NMR(CD₃OD, 400 MHz) δ 7.93 (s, 1H), 7.05-7.12 (m, 9H), 6.85 (d, J =10.0 Hz, 2H), 6.79 (d, J = 8.6 Hz, 2H), 6.47(s, 1H), 3.61 (m, 2H), 3.48(m, 4H), 3.25 (t, J = 7.6 Hz, 2H), 3.01 (t, J = 10.4 Hz, 2H), 2.59 (m,2H), 1.99 (m, 2H), 1.77-1.87 (m, 3H), 1.51-1.61 (m, 3H). MS (ESI) m/z:483 [M + H]⁺. 48 20i

¹H-NMR(CD₃OD, 400 MHz) δ 7.93 (s, 1H), 7.05-7.12 (m, 9H), 6.85 (d, J =10.0 Hz, 2H), 6.79 (d, J = 8.6 Hz, 2H), 6.47(s, 1H), 3.61 (m, 2H), 3.48(m, 4H), 3.25 (t, J = 7.6 Hz, 2H), 3.01 (t, J = 10.4 Hz, 2H), 2.59 (m,2H), 1.99 (m, 2H), 1.77-1.87 (m, 3H), 1.51-1.61 (m, 3H). MS (ESI) m/z:481 [M + H]⁺. 49 20j

¹H-NMR(CD₃OD, 400 MHz) δ 7.15-7.07 (m, 5H), 7.02 (d, J = 8.2 Hz, 2H),6.91 (d, J = 8.1 Hz, 2H), 6.84 (d, J = 8.0 Hz, 2H), 6.77 (d, J = 8.1 Hz,2H), 3.41 (m, 4H), 3.06 (m, 2H), 2.74 (m, 1H), 2.52 (m, 2H), 1.96 (m,2H), 1.76 (m, 2H), 1.54 (m, 2H). MS (ESI) m/z: 414 [M + H]⁺. 50 20k

¹H-NMR(CD₃OD, 400 MHz) δ 7.17 (d, J = 7.8 Hz, 2H), 7.12-7.06 (m, 5H),6.96 (m, 4H), 6.86 (m, 2H), 3.46 (m, 2H), 3.40 (t, J = 6.8 Hz, 2H), 3.10(m, 2H), 2.82 (m, 1H), 2.50 (m, 2H), 2.35 (s, 3H), 2.02 (m, 2H), 1.83(m, 2H), 1.53 (m, 2H). MS (ESI) m/z: 412 [M + H]⁺. 51 20l

¹H-NMR(CD₃OD, 400 MHz) δ 8.26 (d, J = 8.6 Hz, 2H) 7.47 (d, J = 8.6 Hz,2H), 7.21-7.09 (m, 5H), 6.64 (d, J = 8.4 Hz, 2H), 6.30 (d, J = 8.7 Hz,2H), 3.48 (m, 2H), 3.25 (m, 2H), 3.15 (m, 2H), 2.51 (m, 2H), 2.12 (m,4H), 1.59 (m, 4H), 1.15 (m, 2H). MS (ESI) m/z: 484 [M + H]⁺.

[Example 52] Preparation of(E)-4-(5-hydroxy-1-(4-(1-isopropylazetidin-3-yl)phenyl)-2-phenylpent-1-en-1-yl)phenol(22a)

Step 1: Preparation of methyl(E)-5-(4-(1-isopropylazetidin-3-yl)phenyl)-4-phenyl-5-(4-(pivaloyloxy)phenyl)pent-4-enoate(E-2)

Compound E-1 (0.03 g, 0.06 mmol), acetone (0.14 mL, 1.9 mmol), andsodium triacetoxyborohydride (NaBH(OAc)₃, 41 mg, 0.19 mmol) were addedto dichloroethane (3 mL), and stirred at room temperature for 1 hour.Water and ethyl acetate were further added to the reaction solution andan organic layer was extracted. The organic layer was dried withanhydrous Na₂SO₄ and filtered. The solvent was distilled under reducedpressure to obtain a residue, which was purified using columnchromatography, thereby obtaining 18 mg of the desired compound E-2(54%).

Step 2: Preparation of(E)-4-(5-hydroxy-1-(4-(1-isopropylazetidin-3-yl)phenyl)-2-phenylpent-1-en-1-yl)phenol(22a)

4 mg of the desired compound 22a (27%) was obtained by the same processas step 3 of Example 19, using compound E-2.

Examples 53 to 82

Compounds 22b to 22ae were prepared, using the process of Example 52.Identification data of the thus-prepared compounds 22b to 22ae is shownin the following Table 5.

TABLE 5

          Example         Cmpd No.

          R           Identification data 52 22a

¹H-NMR(CD₃OD, 400 MHz) δ 7.17-7.08 (m, 5H), 7.08-7.02 (m, 4H), 6.94 (d,J = 8.2 Hz, 2H), 6.78 (d, J = 8.5 Hz, 2H), 4.38 (t, J = 8.3 Hz, 2H),4.21 (m. 1H), 4.10 (t, J = 9.8 Hz, 2H), 3.98 (m, 1H), 3.43 (t, J = 7.5Hz, 2H), 2.55 (m, 2H), 1.56 (m, 2H), 1.24 (d, J = 6.4 Hz, 6H). MS (ESI)m/z: 428 [M + H]⁺. 53 22b

¹H-NMR(CD₃OD, 400 MHz) δ 7.15-7.07 (m, 5H), 7.05-7.01 (m, 4H), 6.94 (m,2H), 6.76 (d, J = 8.4 Hz, 2H), 4.46 (m, 2H), 4.34 (m, 1H), 4.22 (m, 1H),4.02 (m, 2H), 3.41 (t, J = 6.7 Hz, 2H), 2.91 (s, 3H), 2.53 (m, 2H), 1.54(m, 2H). MS (ESI) m/z: 400 [M + H]⁺. 54 22c

¹H-NMR(CD₃OD, 400 MHz) δ 7.17-7.03 (m, 9H), 6.96 (d, J = 8.1 Hz, 2H),6.78 (d, J = 8.4 Hz, 2H), 4.46 (m, 4H), 4.32 (m, 1H), 3.43 (t, J = 6.7Hz, 2H), 3.35 (s, 3H), 3.21 (s, 3H), 2.55 (m, 2H), 1.55 (m, 2H). MS(ESI) m/z: 415 [M + H]⁺. 55 22d

¹H-NMR(CD₃OD, 400 MHz) δ 7.15-7.09 (m, 5H), 7.06-7.00 (m, 4H), 6.94 (d,J = 8.04, 2H), 6.76 (d, J = 8.6 Hz, 2H), 4.42 (t, J = 9.3 Hz, 2H), 4.32(m, 1H), 4.19 (m, 1H), 3.98 (m, 1H), 3.41 (t, J = 6.7 Hz, 2H), 2.53 (m,2H), 1.54 (m, 2H), 0.86 (m, 4H). MS (ESI) m/z: 426 [M + H]⁺. 56 22e

¹H-NMR(CD₃OD, 400 MHz) δ 7.17-7.07 (m, 9H), 6.98 (d, J = 8.2 Hz, 2H),6.88 (d, J = 8.2 Hz, 2H), 4.07 (t, J = 8.2 Hz, 2H), 3.76 (m, 1H), 3.65(t, J = 8.8 Hz, 2H), 3.39 (t, J = 6.7 Hz, 2H), 3.01 (m, 1H), 2.49 (m,2H), 2.34 (s, 3H), 1.54 (m, 2H), 1.09 (d, J = 6.4 Hz, 6H). MS (ESI) m/z:426 [M + H]⁺. 57 22f

¹H-NMR(CD₃OD, 400 MHz) δ 7.17 (d, J = 7.8 Hz, 2H), 7.12-7.06 (m, 4H),6.97 (m, 2H), 6.85 (m, 2H), 3.58 (m, 2H), 3.40 (t, J = 6.8 Hz, 2H), 3.12(m, 2H), 2.90 (s, 3H), 2.80 (m, 1H), 2.50 (m, 2H), 2.35 (s, 3H), 2.07(m, 2H), 1.88 (m, 2H), 1.53 (m, 2H). MS (ESI) m/z: 426 [M + H]⁺. 58 22g

¹H-NMR(CD₃OD, 400 MHz) δ 7.17-7.07 (m, 5H), 7.02 (d, J = 8.6 Hz, 2H),6.92 (d, J = 8.2 Hz, 2H), 6.85 (d, J = 8.2 Hz, 2H), 6.77 (d, J = 8.6 Hz,2H), 3.48 (m, 3H), 3.41 (t, J = 6.7 Hz, 2H), 3.13 (m, 2H), 2.74 (m, 1H),2.52 (m, 2H), 2.05 (m, 2H), 1.86 (m, 2H), 1.54 (m, 2H), 1.37 (d, J = 6.7Hz, 6H). MS (ESI) m/z: 456 [M + H]⁺. 59 22h

¹H-NMR(CD3OD, 400 MHz) δ 7.19-7.09 (m, 6H), 6.85 (m, 4H), 6.71 (m, 2H),6.64 (m, 1H), 3.77 (m, 2H), 3.58 (m, 3H), 3.41 (t, J = 6.8 Hz, 2H), 3.34(m, 2H), 3.21 (m, 2H), 2.50 (m, 2H), 1.53 (m, 2H), 1.40 (d, J = 6.6 Hz,6H). MS (ESI) m/z: 456 [M + H]+. 60 22i

¹H-NMR(CD3OD, 400 MHz) δ 7.15-7.06 (m, 5H), 7.01 (d, J = 8.4 Hz, 2H),6.90 (d, J = 8.2 Hz, 2H), 6.84 (d, J = 8.2 Hz, 2H), 6.75 (d, J = 8.5 Hz,2H), 3.68 (m, 2H), 3.41 (t, J = 6.6 Hz, 2H), 3.24 (m, 2H), 2.79 (m, 2H),2.52 (m, 2H), 2.02 (m, 2H), 1.78 (m, 2H), 1.54 (m, 2H), 0.97 (m, 4H). MS(ESI) m/z: 454 [M + H]+. 61 22j

¹H-NMR(CD₃OD, 400 MHz) δ 7.17-7.10 (m, 7H), 7.05 (d, J = 8.4 Hz, 2H),6.91 (d, J = 8.2 Hz, 2H), 6.79 (d, J = 8.4 Hz, 2H), 6.06 (s, 1H), 4.02(m, 2H), 3.53 (m, 2H), 3.44 (t, J = 6.7 Hz, 2H), 2.96 (m, 1H), 2.81 (s,2H), 2.55 (m, 2H), 1.56 (m, 2H), 1.03 (m, 4H). MS (ESI) m/z: 452 [M +H]⁺. 62 22k

¹H-NMR(CD₃OD, 400 MHz) δ 7.17-7.10 (m, 7H), 7.05 (d, J = 8.4 Hz, 2H),6.91 (d, J = 8.2 Hz, 2H), 6.80 (d, J = 8.4 Hz, 2H), 6.07 (s, 1H), 3.88(s, 2H), 3.64 (m, 2H), 3.44 (t, J = 6.6 Hz, 2H), 3.23 (m, 1H), 2.81 (m,2H), 2.56 (m, 2H), 1.55 (m, 2H), 1.42 (d, J = 6.6 Hz, 6H). MS (ESI) m/z:454 [M + H]⁺. 63 22l

¹H-NMR(CD₃OD, 400 MHz) δ 7.22-7.12 (m, 8H), 6.92 (d, J = 8.3 Hz, 2H),6.73 (m, 2H), 6.66 (s, 1H), 6.06 (s, 1H), 4.02 (m, 2H), 3.48 (m, 2H),3.43 (t, J = 6.8 Hz, 2H), 2.95 (m, 1H), 2.81 (s, 2H), 2.54 (m, 2H), 1.56(m, 2H), 1.04 (m, 4H). MS (ESI) m/z: 452 [M + H]⁺. 64 22m

¹H-NMR(CD₃OD, 400 MHz) δ 7.22-7.12 (m, 9H), 6.93 (d, J = 6.8 Hz, 2H),6.73 (d, J = 7.6 Hz, 2H), 6.08 (s, 1H), 3.88 (s, 2H), 3.64 (m, 2H), 3.44(t, J = 6.6 Hz, 2H), 3.23 (m, 1H), 2.80 (m, 2H), 2.54 (m, 2H), 1.56 (m,2H), 1.41 (d, J = 6.4 Hz, 6H). MS (ESI) m/z: 454 [M + H]⁺. 65 22n

¹H-NMR(CD₃OD, 400 MHz) δ 7.17-7.07 (m, 6H), 7.03 (m, 2H), 6.77 (m, 2H),6.66 (m, 2H), 3.39 (d, J = 8.6 Hz, 1H), 3.89 (m, 1H), 3.72 (m, 2H), 3.63(m, 2H), 3.41 (t, J = 6.7 Hz, 2H), 3.16 (m, 3H), 2.95 (m, 1H), 2.50 (m,2H), 2.21 (m, 2H), 1.86 (m, 2H), 1.74 (m, 4H), 1.55 (m, 2H). MS (ESI)m/z: 483 [M + H]⁺. 66 22o

¹H-NMR(CD₃OD, 400 MHz) δ 8.14 (d, J = 8.6 Hz, 2H), 7.38 (d, J = 8.6 Hz,2H), 7.07-7.03 (m, 5H), 6.86 (d, J = 8.2 Hz, 2H), 6.76 (d, J = 8.2 Hz,2H), 3.57 (m, 2H), 3.32 (t, J = 6.4 Hz, 2H), 3.14 (m, 2H), 2.68 (m, 2H),2.41 (m, 2H), 1.88 (m, 2H), 1.76 (m, 2H), 1.46 (m, 2H), 0.87 (m, 4H). MS(ESI) m/z: 483 [M + H]⁺. 67 22p

¹H-NMR(CD₃OD, 400 MHz) δ 7.18-7.08 (m, 5H), 7.02 (m, 2H), 6.65 (m, 2H),6.39 (m, 2H), 3.81 (m, 2H), 3.64 (m, 2H), 3.41 (t, J = 6.7 Hz, 2H), 3.23(m, 2H), 2.96 (m, 2H), 2.51 (m, 3H), 2.17 (m, 2H), 1.97 (m, 2H), 1.72(m, 2H), 1.53 (m, 2H), 1.42 (m, 2H), 1.29 (m, 2H). MS (ESI) m/z: 497[M + H]⁺. 68 22q

¹H-NMR(CD₃OD, 400 MHz) δ 7.16-7.07 (m, 5H), 7.02 (d, J = 8.8 Hz, 2H),6.78 (m, 2H), 6.66 (m, 2H), 3.72 (m, 2H), 3.49 (m, 2H), 3.41 (t, J = 6.7Hz, 2H), 2.95 (m, 4H), 2.51 (m, 2H), 2.34 (m, 2H), 2.25 (m, 2H), 1.91(m, 2H), 1.53 (m, 2H). MS (ESI) m/z: 469 [M + H]⁺. 69 22r

¹H-NMR(CD₃OD, 400 MHz) δ 7.21-7.10 (m, 7H), 6.90 (m, 4H), 6.72 (d, J =7.9 Hz, 2H), 3.71 (m, 2H), 3.43 (t, J = 6.8 Hz, 2H), 3.27 (m, 2H), 2.80(m, 2H), 2.52 (m, 2H), 2.01 (m, 2H), 1.79 (m, 2H), 1.52 (m, 2H), 0.98(m, 4H). MS (ESI) m/z: 454 [M + H]⁺. 70 22s

¹H-NMR(CD₃OD, 400 MHz) δ 7.04-6.86 (m, 9H), 6.78 (d, J = 8.1 Hz, 2H),6.65 (d, J = 8.5 Hz, 2H), 6.61-6.58 (m, 2H), 3.61-3.51 (m, 2H),3.36-3.28 (m, 5H), 2.96-2.90 (m, 1H), 2.43- 2.39 (m, 2H), 2.33-2.28 (m,1H), 2.01-1.96 (m, 1H), 1.45-1.41 (m, 2H), 1.26-1.24 (m, 6H). MS (ESI)m/z: 442 [M + H]⁺. 71 22t

¹H-NMR(CD₃OD, 400 MHz) δ 7.18-7.09 (m, 5H), 7.05-7.02 (m, 4H), 6.94-6.89(m, 2H), 6.80-6.78 (m, 2H), 4.28-4.25 (m, 1H), 3.90- 3.73 (m, 3H),3.55-3.51 (m, 1H), 3.45-3.41 (m, 3H), 3.28-3.23 (m, 1H), 3.05-2.96 (m,1H), 2.56-2.52 (m, 2H), 2.44-2.39 (m, 1H), 2.25- 2.19 (m, 1H), 1.59-1.53(m, 2H), 1.00-0.95 (m, 4H). MS (ESI) m/z: 440 [M + H]⁺. 72 22u

¹H-NMR(CD₃OD, 400 MHz) δ 7.08-6.97 (m, 6H), 6.92-6.88 (m, 2H), 6.83-6.79(m, 2H), 6.61-6.58 (m, 2H), 6.53 (s, 1H), 3.59-3.51 (m, 2H), 3.38-3.29(m, 5H), 2.96-2.91 (m, 1H), 2.42-2.38 (m, 2H), 2.34-2.24 (m, 1H), 2.05-1.97 (m, 1H), 1.47-1.39 (m, 2H), 1.26-1.24 (m, 6H). MS (ESI) m/z: 442[M + H]⁺. 73 22v

¹H-NMR(CD₃OD, 400 MHz) δ 7.20-7.09 (m, 6H), 7.04-7.00 (m, 2H), 6.93-6.91(m, 2H), 6.71 (d, J = 7.8 Hz, 2H), 6.55 (s, 1H), 3.91-3.65 (m, 2H),3.54-3.50 (m, 2H), 3.42 (t, J = 6.7 Hz, 2H), 3.27-3.22 (m, 1H),3.01-2.96 (m, 1H), 2.54-2.50 (m, 2H), 2.45-2.42 (m, 1H), 2.23- 2.21 (m,1H), 1.58-1.51 (m, 2H), 0.99-0.90 (m, 4H). MS (ESI) m/z: 440 [M + H]⁺.74 22w

¹H-NMR(CD₃OD, 400 MHz) δ 7.20-7.11 (m, 5H), 7.07 (d, J = 8.2 Hz, 2H),7.01-6.98 (m, 1H), 6.80 (d, J = 8.2 Hz, 2H), 6.76-6.70 (m, 1H),6.62-6.53 (m, 2H), 3.54-3.42 (m, 7H), 3.22-3.13 (m, 2H), 2.96-2.90 (m,2H), 2.57- 2.54 (m, 2H), 1.61-1.55 (m, 2H), 1.40 (d, J = 6.3 Hz, 6H). MS(ESI) m/z: 457 [M + H]⁺. 75 22x

¹H-NMR(CD₃OD, 400 MHz) δ 7.21-7.11 (m, 6H), 7.04-7.00 (m, 1H), 6.75-6.69(m, 4H), 6.62-6.57 (m, 2H), 3.58-3.42 (m, 7H), 3.20- 3.15 (m, 2H),2.97-2.92 (m, 2H), 2.56-2.52 (m, 2H), 1.61-1.53 (m, 2H), 1.41 (d, J =6.6 Hz, 6H). MS (ESI) m/z: 457 [M + H]⁺. 76 22y

¹H-NMR(CD₃OD, 400 MHz) δ 7.17-6.98 (m, 8H), 6.89 (d, J = 7.6 Hz, 1H),6.79-6.77 (m, 4H), 3.54-3.32 (m, 5H), 3.12-3.06 (m, 2H), 2.66-2.60 (m,1H), 2.59-2.54 (m, 2H), 1.87- 1.84 (m, 2H), 1.77-1.71 (m, 2H), 1.60-1.53(m, 2H), 1.37 (d, J = 6.5 Hz, 6H). MS (ESI) m/z: 456 [M + H]⁺. 77 22z

¹H-NMR(CD₃OD, 400 MHz) δ 7.17-6.99 (m, 8H), 6.91-6.88 (m, 1H), 6.80-6.78(m, 4H), 3.68-3.64 (m, 2H), 3.44 (dd, J = 6.7 Hz, 2H), 3.26-3.20 (m,2H), 2.81-2.77 (m, 1H), 2.69- 2.62 (m, 1H), 2.58-2.54 (m, 2H), 1.86-1.82(m, 2H), 1.68-1.59 (m, 2H), 1.58-1.53 (m, 2H), 1.00-0.98 (m, 4H). MS(ESI) m/z: 454 [M + H]⁺. 78 22aa

¹H-NMR(CD₃OD, 400 MHz) δ 7.22-7.09 (m, 6H), 7.05-7.00 (m, 1H), 6.93-6.90(m, 1H), 6.83-6.80 (m, 2H), 6.75-6.71 (m, 2H), 6.67- 6.65 (m, 1H),3.49-3.42 (m, 5H), 3.15-3.06 (m, 2H), 2.67-2.60 (m, 1H), 2.56-2.53 (m,2H), 1.89-1.85 (m, 2H), 1.79-1.67 (m, 2H), 1.60- 1.53 (m, 2H), 1.38 (d,J = 7.7 Hz, 6H). MS (ESI) m/z: 456 [M + H]⁺. 79 22ab

¹H-NMR(CD₃OD, 400 MHz) δ 7.21-7.09 (m, 6H), 7.01 (dd, J = 7.9 Hz, 1H),6.90 (d, J = 7.7 Hz, 1H), 6.81-6.80 (m, 1H), 6.74-6.71 (m, 2H),6.68-6.67 (m, 2H), 3.67-3.64 (m, 2H), 3.44 (dd, J = 6.8 Hz, 2H),3.26-3.20 (m, 2H), 2.81-2.78 (m, 1H), 2.69-2.63 (m, 1H), 2.56-2.53 (m,2H), 1.84-1.81 (m, 2H), 1.75-1.65 (m, 2H), 1.59- 1.53 (m, 2H), 1.04-0.95(m, 4H). MS (ESI) m/z: 454 [M + H]⁺. 80 22ac

¹H-NMR(CD₃OD, 400 MHz) δ 7.15-7.07 (m, 6H), 6.93-6.85 (m, 4H), 6.70 (m,2H), 6.63 (s, 1H), 3.53 (m, 2H), 3.41 (t, J = 6.8 Hz, 2H), 3.08 (m, 2H),2.87 (s, 3H), 2.72 (m, 1H), 2.51 (m, 2H), 2.02 (m, 2H), 1.86 (m, 2H),1.56 (m, 2H). MS (ESI) m/z: 428 [M + H]⁺. 81 22ad

¹H-NMR(CD₃OD, 400 MHz) δ 7.21-7.11 (m, 7H), 6.85 (m, 2H), 6.71 (m, 4H),3.70 (m, 4H), 3.42 (t, J = 6.7 Hz, 2H), 3.18 (m, 3H), 2.53 (m, 2H), 1.53(m, 2H), 1.00 (m, 4H). MS (ESI) m/z: 455 [M + H]⁺. 82 22ae

¹H-NMR(CD₃OD, 400 MHz) δ 7.19-7.09 (m, 6H), 6.97-6.85 (m, 4H), 6.69 (m,2H), 6.63 (s, 1H), 3.60 (m, 2H), 3.41 (t, J = 6.7 Hz, 2H), 3.16 (m, 2H),3.00 (m, 2H), 2.74 (m, 1H), 2.51 (m, 2H), 2.02 (m, 2H), 1.85 (m, 2H),1.52 (m, 2H), 1.36 (m, 3H). MS (ESI) m/z: 442 [M + H]⁺.

[Example 83] Preparation of(Z)-5-(4-aminophenyl)-5-(4-(4-methylpiperazin-1-yl)phenyl)-4-phenylpent-4-en-1-ol(26a)

Compound F-1 (0.01 g, 0.02 mmol) and ammonium chloride (11 mg, 0.21mmol) were added to methanol (0.5 mL) and tetrahydrofuran (0.5 mL), thetemperature was lowered to 0° C., and zinc (13 mg, 0.21 mmol) was addedthereto Stirring was performed at room temperature for 12 hours.Filtration was performed using celite, and the solvent was distilledunder reduced pressure to obtain a residue, which was purified usingcolumn chromatography, thereby obtaining 9 mg of the desired compound26a (99%).

Examples 84 to 86

Compounds 26b to 26d were prepared, using the process of Example 83.Identification data of the thus-prepared compounds 26a to 26d is shownin the following Table 6.

TABLE 6

Cmpd Example No. R Identification data 83 26a

¹H-NMR(CD₃OD, 400 MHz) δ 7.43 (s, 4H), 7.21-7.14 (m, 5H), 6.82 (d, J =8.8 Hz, 2H, 6.71 (d, J = 8.8 Hz, 2H). 3.70 (m, 2H), 3.55 (m, 2H), 3.44(m, 2H), 3.17 (m, 2H), 3.14 (m, 5H), 2.49 (m, 2H), 1.55 (m, 2H). MS(ESI) m/z: 428 [M + H]⁺. 84 26b

¹H-NMR(CD₃OD, 400 MHz) δ 7.42 (m, 2H), 7.25-7.09 (m, 7H), 7.05 (m, 2H),6.82 (m, 2H), 3.96 (m, 2H), 3.79 (m, 2H), 3.61 (m, 2H), 3.42 (m, 3H),3.17 (m, 2H), 2.47 (m, 2H), 1.54 (m, 2H), 1.41 (d, J = 6.6 Hz, 6H). MS(ESI) m/z: 456 [M + H]⁺. 85 26c

¹H-NMR(CD₃OD), 400 MHz) δ 7.44 (m, 4H), 7.25-7.11 (m, 5H), 6.97 (m, 2H),6.88 (m, 2H), 3.59 (m, 4H), 3.42 (t, J = 6.6 Hz, 2H), 3.14 (m, 2H), 2.52(m, 2H), 2.01 (m, 4H), 1.56 (m, 2H), 1.39 (d, J = 6.6 Hz, 6H). MS (ESI)m/z: 455 [M + H]⁺. 86 26d

¹H-NMR(CD₃OD, 400 MHz) δ 7.32 (s, 4H), 7.08-7.01 (m, 5H), 6.85 (d, J =8.1 Hz 2H), 6.76 (d, J = 8.1 Hz, 2H), 3.59 (m, 2H), 3.30 (t, J = 6.5 Hz,2H), 3.15 (m, 2H), 2.69 (m, 2H), 2.38 (m, 2H), 1.88 (m, 2H), 1.78 (m,2H), 1.44 (m, 2H), 0.86 (m, 4H). MS (ESI) m/z: 453 [M + H]⁺.

[Example 87] Preparation of(E)-N-(4-(5-hydroxy-1-(4-(4-isopropylpiperazin-1-yl)phenyl)-2-phenylpent-1-en-1-yl)phenyl)methanesulfonamide(27a)

Compound 26b (5 mg, 10 μmol) and triethylamine (3 μL, 0.02 mmol) wereadded to dichloromethane (2 mL), the temperature was lowered to 0° C.,and methanesulfonyl chloride (1 μL, 0.01 mmol) was added thereto.Stirring was performed at room temperature for 12 hours. Saturatedsodium hydrogen carbonate and dichloromethane were further added to thereaction solution and an organic layer was extracted. The organic layerwas dried with anhydrous Na₂SO₄ and filtered. The solvent was distilledunder reduced pressure to obtain a residue, which was purified usingcolumn chromatography and then dissolved in methanol:dichloromethane(1:1), the temperature was lowered to 0° C., a 1M aqueous HCl solutionwas slowly added thereto, and distillation under reduced pressure wasperformed, thereby obtaining 1 mg of the desired compound 27a (17%).

Examples 87 to 94

Compounds 27b to 27h were prepared, using the process of Example 87.Identification data of the thus-prepared compounds 27a to 27h is shownin the following Table 7.

TABLE 7

R^(a) R^(b) R^(c) R^(d)

87 27a

H H

¹H-NMR(CD₃OD, 400 MHz) δ 7.43 (s, 4H), 7.19-7.13 (m, 5H), 6.82 (d, J =7.6 Hz, 2H), 6.71 (d, J = 8.1 Hz 2H), 4.10 (t, J = 5.8 Hz, 2H), 3.76 (m,2H), 3.54 (m, 3H), 3.21 (m, 2H), 2.98 (m, 5H), 2.56 (m, 2H), 1.74 (m,2H), 1.39 (d, J = 6.5 Hz, 6H). MS (ESI) m/z: 534 [M + H]⁺. 88 27b

H

¹H-NMR(CD₃OD, 400 MHz) δ 7.26 (d, J = 8.4 Hz, 2H), 7.21-7.14 (m, 7H),6.82 (d, J = 8.6 Hz, 2H), 6.70 (d, J = 7.5 Hz, 2H), 4.10 (t, J = 5.8 Hz,2H), 3.77 (m, 2H), 3.54 (m, 3H), 3.21 (m, 2H), 2.96 (m, 8H), 2.59 (m,2H), 1.73 (m, 2H), 1.39 (d, J = 6.6 Hz, 6H). MS (ESI) m/z: 612 [M + H]⁺.89 27c

H H

¹H-NMR(CD₃OD, 400 MHz) δ 7.43 (d, J = 8.5 Hz, 2H), 7.07-6.98 (m, 7H),6.82 (d, J = 8.2 Hz, 2H), 6.75 (d, J = 8.2 Hz, 2H), 3.57 (m, 2H), 3.32(t, J = 6.7 Hz, 2H), 3.14 (m, 2H), 2.70 (m, 2H), 2.41 (m, 2H), 2.03 (s,3H), 1.92 (m, 2H), 1.68 (m, 2H), 1.44 (m, 2H), 0.83 (m, 4H). MS (ESI)m/z: 495 [M + H]⁺. 90 27d

H

¹H-NMR(CD₃OD), 400 MHz) δ 7.45 (d, J = 8.5 Hz, 2H), 7.06-7.01 (m, 7H),6.82 (d, J = 8.3 Hz, 2H), 6.76 (d, J = 8.3 Hz, 2H), 3.84 (t, J = 6.3 Hz,2H), 6.70 (m, 2H), 3.13 (m, 2H), 2.67 (m, 2H), 2.43 (m, 2H), 2.03 (s,3H), 1.92 (m, 2H), 1.83 (s, 3H), 1.72 (m, 2H), 1.51 (m, 2H), 0.84 (m,4H). MS (ESI) m/z: 537 [M + H]⁺. 91 27e

H H

¹H-NMR(CD₃OD, 400 MHz) δ 7.27 (d, J = 8.5 Hz, 2H), 7.19-7.09 (m, 7H),6.81 (d, J = 8.6 Hz, 2H), 6.68 (d, J = 8.7 Hz, 2H), 3.76 (m, 2H), 3.51(m, 2H), 3.41 (t, J = 6.5 Hz, 2H), 3.21 (m, 2H), 2.92 (m, 2H), 2.57 (m,1H), 2.50 (m, 2H), 1.52 (m, 2H), 1.39 (d, J = 6.6 Hz, 6H), 1.29 (s, 1H),1.04 (m, 2H), 0.96 (m, 2H). MS (ESI) m/z: 560 [M + H]⁺. 92 27f

H H

¹H-NMR(CD₃OD, 400 MHz) δ 7.33 (m, 4H), 7.19-7.12 (m, 5H), 6.83 (d, J =8.7 Hz, 2H), 6.70 (d, J = 8.8 Hz, 2H), 4.11 (t, J = 6.0 Hz, 2H), 3.77(m, 2H), 3.53 (m, 3H), 3.20 (m, 2H), 2.95 (m, 5H), 2.58 (m, 2H), 1.73(m, 2H), 1.38 (d, J = 6.6 Hz, 6H). MS (ESI) m/z: 498 [M + H]⁺. 93 27g

H H

¹H-NMR(CD₃OD, 400 MHz) δ 7.27 (d, J = 8.4 Hz, 2H), 7.19-7.09 (m, 7H),6.90 (m, 4H), 3.67 (m, 3H), 3.41 (t, J = 6.4 Hz, 2H), 2.77 (m, 2H), 2.55(m, 3H), 2.02 (m, 2H), 1.76 (m, 2H), 1.54 (m, 2H), 0.96 (m, 4H). MS(ESI) m/z: 557 [M + H]⁺. 94 27h

H H

¹H-NMR(CD₃OD, 400 MHz) δ 7.28-7.11 (m, 9H), 6.96 (m 4H), 3.73 (m, 4H),3.50 (m, 2H), 2.83 (m, 2H), 2.54 (m, 2H), 2.18 (m, 1H), 1.98 (m, 1H),1.54(m, 2H), 0.95 (m, 4H). MS (ESI) m/z: 531 [M + H]⁺.

[Example 95] Preparation of(E)-4-(5-hydroxy-1-(4-(4-isopropylpiperazin-1-yl)phenyl)-2-phenylpent-1-en-1-yl)phenylmethanesulfonate (28a)

Compound G-1 (10 mg, 22 μmol) and diisopropylethylamine (8 μL, 0.04mmol) were added to dichloromethane (2 mL), the temperature was loweredto 0° C., and methanesulfonyl chloride (3 μL, 0.02 mmol) was addedthereto. Stirring was performed at room temperature for 12 hours.Saturated sodium hydrogen carbonate and dichloromethane were furtheradded to the reaction solution and an organic layer was extracted. Theorganic layer was dried with anhydrous Na₂SO₄ and filtered. The solventwas distilled under reduced pressure to obtain a residue, which waspurified using column chromatography, thereby obtaining 1 mg of thedesired compound 28a (9%).

Examples 96 to 101

Compounds 28b to 28g were prepared, using the process of Example 95.Identification data of the thus-prepared compounds 28a to 28g is shownin the following Table 8.

TABLE 8

Cmpd Example No. R^(a) R^(b) R^(c) Identification data 95 28a

H

¹H-NMR(CD₃OD, 400 MHz) δ 7.32 (m, 4H), 7.19- 7.10 (m, 5H), 6.81 (d, J =8.7 Hz, 2H), 6.70 (d, J = 8.8 Hz, 2H), 3.77 (m, 2H), 3.53 (m, 3H), 3.41(t, J = 6.6 Hz, 2H), 3.25 (s, 3H), 3.20 (m, 2H), 2.92 (m, 2H), 2.49 (m,2H), 1.54 (m, 2H), 1.38 (d, J = 6.6 Hz, 6H). MS (ESI) m/z: 535 [M + H]⁺.96 28b

¹H-NMR(CD₃OD, 400 MHz) δ 7.45 (d, J = 8.5 Hz, 2H), 7.06-6.99 (m, 7H),6.82 (d, J = 8.3 Hz, 2H), 6.76 (d, J = 8.3 Hz, 2H), 3.84 (t, J = 6.3 Hz,2H), 3.59 (m, 2H), 3.14 (m, 2H), 2.68 (m, 2H), 2.43 (m, 2H), 2.03 (s,3H), 1.92 (m, 2H), 1.83 (s, 3H), 1.55 (m, 2H), 1.52 (m, 2H), 0.84 (m,4H). MS (ESI) m/z: 613 [M + H]⁺. 97 28c

H

¹H-NMR(CD₃OD, 400 MHz) δ 7.35-7.29 (m, 4H), 7.19-7.10 (m, 5H), 6.81 (d,J = 8.8 Hz, 2H), 6.71 (d, J = 8.8 Hz, 2H), 3.77 (m, 2H), 3.54 (m, 3H),3.41 (t, J = 6.6 Hz, 2H), 3.21 (m, 2H), 2.97 (m, 2H), 2.82 (m, 1H), 2.49(m, 2H), 1.53 (m, 2H), 1.38 (d, J = 6.6 Hz, 6H), 1.14 (m, 4H). MS (ESI)m/z: 564 [M + H]⁺. 98 28d

H

¹H-NMR(CD₃OD), 400 MHz) δ 7.38 (m, 4H), 7.18- 7.11 (m, 5H), 6.81 (d, J =8.8 Hz, 2H), 6.70 (d, J = 8.8 Hz, 2H), 3.77 (m, 2H), 3.51 (m, 3H), 3.41(t, J = 6.4 Hz, 2H), 3.21 (m, 2H), 2.92 (m, 2H), 2.48 (m, 2H), 1.53 (m,2H), 1.38 (d, J = 6.6 Hz, 6H). MS (ESI) m/z 589 [M + H]⁺. 99 28e

H

¹H-NMR(CD₃OD, 400 MHz) δ 7.25 (d, J = 8.6 Hz, 2H), 7.15 (m, 5H), 7.05(d, J = 8.5 Hz, 2H), 6.81 (d, J = 8.8 Hz, 2H), 6.68 (d, J = 8.8 Hz, 2H),3.72 (m, 2H), 3.52 (m, 2H), 3.39 (t, J = 6.7 Hz, 2H), 3.18 (m, 2H), 2.92(m, 5H), 2.51 (m, 2H), 1.54 (m, 2H), 1.36 (s, 9H). MS (ESI) m/z: 513[M + H]⁺. 100 28f H

¹H-NMR(CD₃OD, 400 MHz) δ 7.20-7.16 (m, 5H), 7.12 (d, J = 7.2 Hz, 2H),7.04 (d, J = 8.7 Hz, 2H), 6.68 (d, J = 8.7 Hz, 2H), 3.89 (m, 2H), 3.62(m, 2H), 3.48 (m, 2H), 3.42 (t, J = 6.7 Hz, 2H), 3.07 (m, 2H), 2.98 (s,3H), 2.54 (m, 2H), 1.56 (m, 2H), 1.28 (s, 9H). MS (ESI) m/z: 513 [M +H]⁺. 101 28g

¹H-NMR(CD₃OD, 400 MHz) δ 7.24 (m, 2H), 7.18- 7.06 (m, 7H), 6.82 (d, J =6.9 Hz, 2H), 6.69 (d, J = 8.8 Hz, 2H), 3.93 (t, J = 6.1 Hz, 2H), 3.72(m, 2H), 3.52 (m, 2H), 3.19 (m, 2H), 2.95 (m, 5H), 2.52 (m, 2H), 1.63(m, 2H), 1.37 (s, 9H), 1.08 (s, 9H). MS (ESI) m/z: 597 [M + H]⁺.

[Example 102] Preparation of(E)-4-(5-hydroxy-1-(4-((4-methylpiperazin-1-yl)methyl)phenyl)-2-phenylpent-1-en-1-yl)phenol(30a)

Step 1: Preparation of methyl(E)-5-(4-((4-methylpiperazin-1-yl)methyl)phenyl)-4-phenyl-5-(4-(pivaloyloxy)phenyl)pent-4-enoate(H-2)

Compound H-1 (0.01 g, 0.02 mmol), 1-methylpiperizine (7 μL, 0.06 mmol),and sodium triacetoxyborohydride (NaBH(OAc)₃, 14 mg, 0.06 mmol) wereadded to dichloroethane (3 mL), and heated at 50° C. for 12 hour. Waterand ethyl acetate were further added to the reaction solution and anorganic layer was extracted. The organic layer was dried with anhydrousNa₂SO₄ and filtered. The solvent was distilled under reduced pressure toobtain a residue, which was purified using column chromatography,thereby obtaining 12 mg of the desired compound H-2 (99%).

Step 2: Preparation of(E)-4-(5-hydroxy-1-(4-((4-methylpiperazin-1-yl)methyl)phenyl)-2-phenylpent-1-en-1-yl)phenol(30a)

2 mg of the desired compound 30a (18%) was obtained by the same processas step 3 of Example 19, using compound H-2.

Example 103

Compound 30b was prepared, using the process of Example 102.Identification data of the thus-prepared compounds 30a and 30b is shownin the following Table 9.

TABLE 9

Cmpd Example No. R Identification data 102 30a OH ¹H-NMR(CD₃OD, 400MHz), δ 7.19 (m, 2H), 7.16-7.10 (m, 5H), 7.07 (d, J = 8.4 Hz, 2H), 7.02(d, J = 7.8 Hz, 2H), 6.80 (d, J = 8.3 Hz, 2H), 4.17 (s, 2H), 3.55 (m,8H), 3.43 (t, J = 6.7 Hz, 2H), 2.96 (s, 3H), 2.55 (m, 2H), 1.56 (m, 2H).MS (ESI) m/z: 443 [M + H]⁺. 103 30b NO₂ ¹H-NMR(CD₃OD, 400 MHz) δ 8.26(d, J = 7.9 Hz, 2H), 7.53 (d, J = 8.1 Hz, 2H), 7.31 (m, 2H), 7.17 (m,5H), 7.06 (d, J = 7.2 Hz, 2H), 4.32 (s, 2H), 3.61 (m, 8H), 3.43 (t, J =6.2 Hz, 2H), 2.98 (s, 3H), 2.53 (m, 2H), 1.56 (m, 2H), MS (ESI) m/z: 472[M + H]⁺.

[Example 104] Preparation of(Z)-4-(5-hydroxy-2-phenyl-1-(1-(2-(pyrrolidin-1-yl)ethyl)indolin-5-yl)pent-1-en-1-yl)phenol(33)

Step 1: Preparation of methyl(Z)-5-(indolin-5-yl)-4-phenyl-5-(4-(pivaloyloxy)phenyl)pent-4-enoate(I-2)

0.2 g of the desired compound I-2 (99%) was obtained by the same processas step 1 of Example 40, using compound I-1.

Step 2: Preparation of methyl(Z)-4-phenyl-5-(4-(pivaloyloxy)phenyl)-5-(1-(2-(pyrrolidin-1-yl)ethyl)indolin-5-yl)pent-4-enoate(I-3)

Compound I-2 (20 mg, 0.04 mmol), potassium carbonate (17 mg, 0.12 mmol),and sodium iodide (0.06 mg, 0.414 μmol) were added to dimethylformamide(1 mL), and stirred at room temperature for 12 hour. Saturated sodiumhydrogen carbonate and ethyl acetate were added to the reaction solutionand an organic layer was extracted. The organic layer was dried withanhydrous Na₂SO₄ and filtered. The solvent was distilled under reducedpressure to obtain a residue, which was purified using columnchromatography, thereby obtaining 3 mg of the desired compound I-3(11%).

Step 3: Preparation of(Z)-4-(5-hydroxy-2-phenyl-1-(1-(2-(pyrrolidin-1-yl)ethyl)indolin-5-yl)pent-1-en-1-yl)phenol(33)

1 mg of the desired compound 33 (55%) was obtained by the same processas step 5 of Example 1, using compound I-3.

¹H-NMR (CD₃OD, 400 MHz) δ 7.19-6.97 (m, 7H), 6.77 (d, J=8.5 Hz, 2H),6.64 (m, 2H), 6.41 (d, J=8.7 Hz, 2H), 3.67 (m, 2H), 3.41 (m, 2H), 3.51(m, 2H), 2.06 (m, 6H), 1.55 (m, 2H), 0.89 (m, 4H). MS (ESI) m/z: 469[M+H]⁺.

[Example 105] Preparation of(Z)-4-(5-hydroxy-2-phenyl-1-(1-(2-(piperidin-1-yl)ethyl)-1H-indol-5-yl)pent-1-en-1-yl)phenol(36)

Step 1: Preparation of methyl(Z)-5-(1H-indol-5-yl)-4-phenyl-5-(4-(pivaloyloxy)phenyl)pent-4-enoate(J-2)

24 mg of the desired compound J-2 (99%) was obtained by the same processas step 2 of Example 40, using compound J-1.

Step 2: Preparation of methyl(Z)-4-phenyl-5-(1-(2-(piperidin-1-yl)ethyl)-1H-indol-5-yl)-5-(4-(pivaloyloxy)phenyl)pent-4-enoate(J-3)

9 mg of the desired compound J-3 (31%) was obtained by the same processas step 1 of Example 104, using compound J-2.

Step 3: Preparation of(Z)-4-(5-hydroxy-2-phenyl-1-(1-(2-(piperidin-1-yl)ethyl)-1H-indol-5-yl)pent-1-en-1-yl)phenol(36)

2 mg of the desired compound 36 (18%) was obtained by the same processas step 5 of Example 1, using compound J-3.

¹H-NMR (CD₃OD, 400 MHz) δ 7.30-6.95 (m, 14H), 4.41 (m, 2H), 3.61 (m,4H), 3.44 (m, 2H), 3.11 (m, 2H), 2.56 (m, 2H), 1.93 (m, 6H), 1.57 (m,2H). MS (ESI) m/z: 481 [M+H]⁺.

[Example 106] Preparation of(Z)-4-(1-(4-(2-(3-azabicyclo[3.1.0]hexan-3-yl)ethoxy)phenyl)-5-hydroxy-2-phenylpent-1-en-1-yl)phenol(38a)

Step 1: Preparation of methyl(E)-5-(4-(2-(3-azabicyclo[3.1.0]hexan-3-yl)ethoxy)phenyl)-4-phenyl-5-(4-(pivaloyloxy)phenyl)pent-4-enoate(K-2)

Compound K-1 (10 mg, 0.02 mmol), 3-azabicyclo[3,1,0]hexane (7 mg, 0.06mmol), sodium iodide (0.3 mg, 2 μmol), and triethylamine (11 μL, 0.08mmol) were added to dimethylformamide (1 mL), and heated at 80° C. for12 hours. The solvent was distilled under reduced pressure to obtain aresidue, which was purified using column chromatography, therebyobtaining 6 mg of the desired compound K-2 (53%).

Step 2: Preparation of (Z)-4 (1-(4 (2 (3azabicyclo[3.1.0]hexan-3-yl)ethoxy)phenyl)-5-hydroxy-2-phenylpent-1-en-1-yl)phenol(38a)

3 mg of the desired compound 38a (62%) was obtained by the same processas step 5 of Example 1, using compound K-2.

Examples 107 to 109

Compounds 38b to 38d were prepared, using the process of Example 106.Identification data of the thus-prepared compounds 38a to 38d is shownin the following Table 10.

TABLE 10

Cmpd Example No. R Identification data 106 38a

¹H-NMR(CD₃OD, 400 MHz) δ 7.16-7.07 (m, 5H), 7.02 (d, J = 8.4 Hz, 2H),6.83 (d, J = 8.7 Hz, 2H), 6.76 (d, J = 2H), 6.65 (m, 2H), 4.17 (t, J =4.4 Hz, 2H), 3.70 (m, 2H), 3.57 (t, J = 4.6 Hz, 2H), 3.46 (m, 2H), 3.41(t, J = 6.7 Hz, 2H), 2.52 (m, 2H), 1.86 (m, 2H), 1.54 (m, 2H), 0.84 (m,1H), 0.63 (m, 1H). MS (ESI) m/z: 456 [M + H]⁺. 107 38b

¹H-NMR(CD₃OD, 400 MHz) δ 7.16-7.07 (m, 5H), 7.02 (d, J = 8.5 Hz, 2H),6.83 (d, J = 8.7 Hz, 2H), 6.76 (d, J = 8.5 Hz, 2H), 6.66 (d, J = 8.8 Hz,2H), 4.19 (t, J = 4.7 Hz, 2H), 3.72 (m, 1H), 3.60 (m, 2H), 3.50 (m, 1H),3.41 (t, J = 6.8 Hz, 2H), 3.15 (m, 1H), 2.52 (m, 2H), 2.06 (m, 1H), 1.92(m, 1H), 1.69 (m, 9H), 1.54 (m, 2H). MS (ESI) m/z: 498 [M + H]⁺. 108 38c

¹H-NMR(CD₃OD, 400 MHz) δ 7.93 (s, 1H), 7.13-7.15 (m, 5H), 7.05 (d, J =8.8 Hz, 2H), 6.85 (d, J = 8.8 Hz, 2H), 6.79 (d, J = 8.8 Hz, 2H), 6.68(d, J = 8.8 Hz, 2H), 4.28 (t, J = 10.0 Hz, 2H), 3.90 (m, 2H), 3.75 (m,4H), 3.50 (m, 1H), 3.44 (t, J = 6.8 Hz, 2H), 2.55 (m, 2H), 1.90-2.22 (m,4H), 1.57 (m, 2H). MS (ESI) m/z: 474 [M + H]⁺. 109 38d

¹H-NMR(CD₃OD, 400 MHz) δ 7.93 (s, 1H), 7.13-7.22 (m, 5H), 7.04 (d, J =8.6 Hz, 2H), 6.85 (d, J = 8.6 Hz, 2H), 6.79 (d, J = 8.6 Hz, 2H), 6.67(d, J = 8.8 Hz, 2H), 4.23 (m, 1H), 4.08-4.16 (m, 3H), 3.93 (m, 1H), 3.81(m, 1H), 3.63 (m, 1H), 3.44 (t, J = 6.8 Hz, 3H), 2.55 (m, 2H), 1.92 (m,1H), 1.79 (m, 1H), 1.59 (m, 2H), 0.93 (m, 2H). MS (ESI) m/z: 486 [M +H]⁺.

[Example 110] Preparation of(Z)-4-(1-(4-(2-(2,7-diazaspiro[4.4]nonan-2-yl)ethoxy)phenyl)-5-hydroxy-2-phenylpent-1-en-1-yl)phenol(39)

0.8 mg of the desired compound 39 (24%) was obtained by the same processas step 1 of Example 40, using compound Q.

¹H-NMR (CD₃OD, 400 MHz) δ 7.18-7.07 (m, 5H), 7.0 (d, J=7.0 Hz, 2H), 6.83(d, J=7.2 Hz, 2H), 6.76 (d, J=7.0 Hz, 2H), 6.68 (d, J=7.6 Hz, 2H), 4.23(s, 2H), 3.81 (m, 2H), 3.66 (m, 2H), 3.41 (m, 8H), 2.51 (m, 2H), 2.21(m, 4H), 1.54 (m, 2H). MS (ESI) m/z: 499 [M+H]⁺.

[Example 111] Preparation of 4-((Z)-1-(4-(2-((3aR,6aS)-hexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)ethoxy)phenyl)-5-hydroxy-2-phenylpent-1-en-1-yl)phenol(40)

2 mg of the desired compound 40 (19%) was obtained by the same processas step 1 of Example 40, using compound R.

¹H-NMR (CD₃OD, 400 MHz) δ 7.08-7.16 (m, 5H), 7.04 (d, J=8.8 Hz, 2H),6.85 (d, J=8.0 Hz, 2H), 6.79 (d, J=8.8 Hz, 2H), 6.70 (d, J=8.0 Hz, 2H),4.31 (m, 2H), 4.04 (m, 2H), 3.83 (m, 1H), 3.68 (m, 3H), 3.36-3.49 (m,8H), 2.55 (m, 2H), 1.57 (m, 2H). MS (ESI) m/z: 485 [M+H]⁺.

[Example 112] Preparation of(Z)-4-(1-(4-(dimethyl((4-methylpiperazin-1-yl)methyl)silyl)phenyl)-5-hydroxy-2-phenylpent-1-en-1-yl)phenol(42a)

Step 1: Preparation of methyl(Z)-5-(4-(dimethyl((4-methylpiperazin-1-yl)methyl)silyl)phenyl)-4-phenyl-5-(4-(pivaloyloxy)phenyl)pent-4-enoate(L-2)

8 mg of the desired compound I-2 (34%) was obtained by the same processas step 1 of Example 106, using compound L-1.

Step 2: Preparation of(Z)-4-(1-(4-(dimethyl((4-methylpiperazin-1-yl)methyl)silyl)phenyl)-5-hydroxy-2-phenylpent-1-en-1-yl)phenol(42a)

3 mg of the desired compound 42a (44%) was obtained by the same processas step 5 of Example 1, using compound L-2.

Examples 113 to 116

Compounds 42b to 42e were prepared, using the process of Example 112.Identification data of the thus-prepared compounds 42a to 42e is shownin the following Table 11.

TABLE 11

Cmpd Example No. R Identification data 112 42a

¹H-NMR(CD₃OD, 400 MHz) δ 7.34 (d, J = 7.6 Hz, 2H), 7.20- 7.13 (m, 5H),7.05 (d, J = 8.4 Hz, 2H), 7.00 (d, J = 7.6 Hz, 2H), 6.79 (d, J = 8.4 Hz,2H), 3.67 (m, 8H), 3.43 (t, J = 6.7 Hz, 2H), 3.01 (m, 5H), 2.55 (m, 2H),1.56 (m, 2H), 0.52 (s, 6H). MS (ESI) m/z: 501 [M + H]⁺. 113 42b

¹H-NMR(CD₃OD, 400 MHz) δ 7.29 (m, 2H), 7.12 (m, 5H), 7.00 (m, 4H), 6.77(m, 2H), 3.42 (m, 2H), 3.25 (m, 2H), 2.83 (m, 4H), 2.54 (m, 2H), 1.77(m, 4H), 1.55 (m, 2H). MS (ESI) m/z: 486 [M + H]⁺. 114 42c

¹H-NMR(CD₃OD, 400 MHz) δ 7.31 (m, 2H), 7.15-6.99 (m, 9H), 6.79 (d, J =8.6 Hz, 2H), 3.44 (m, 4H), 2.93 (s, 2H), 2.83 (m, 2H), 2.56 (m, 2H),2.06 (m, 2H), 1.93 (m, 2H), 1.57 (m, 2H), 0.44 (s, 6H). MS (ESI) m/z:472 [M + H]⁺. 115 42d

¹H-NMR(CD₃OD, 400 MHz) δ 7.34 (d, J = 8.0 Hz, 2H), 7.17- 7.12 (m, 5H),7.05 (m, 2H), 7.00 (d, J = 8.0 Hz, 2H), 6.79 (d, J = 8.5 Hz, 2H), 3.56(s, 8H), 3.43 (t, J = 6.7 Hz, 2H), 3.02 (s, 2H), 2.55 (m, 2H), 1.56 (m,2H), 0.52 (s, 6H). MS (ESI) m/z: 487 [M + H]⁺. 116 42e

¹H-NMR(CD₃OD, 400 MHz) δ 7.30 (m, 2H), 7.13 (m, 5H), 7.01 (m, 4H), 6.77(m, 2H), 4.26 (t, J = 6.2 Hz, 2H), 3.74 (m, 2H), 3.52 (m, 2H), 3.40 (m,2H), 3.25 (m, 2H), 3.12 (m, 2H), 2.99 (s, 2H), 2.60 (m, 2H), 1.75 (m,2H), 1.54 (m, 2H), 0.44 (s, 6H). MS (ESI) m/z: 513 [M + H]⁺.

[Example 117] Preparation of(E)-N-(4-(5-hydroxy-1-(4-hydroxyphenyl)-2-phenylpent-1-en-1-yl)phenyl)-2-(piperidin-1-yl)acetamide(44)

Step 1: Preparation of methyl(E)-4-phenyl-5-(4-(2-(piperidin-1-yl)acetamido)phenyl)-5-(4-(pivaloyloxy)phenyl)pent-4-enoate(M-2)

7 mg of the desired compound M-2 (80%) was obtained by the same processas step 1 of Example 106, using compound M-1.

Step 2: Preparation of(E)-N-(4-(5-hydroxy-1-(4-hydroxyphenyl)-2-phenylpent-1-en-1-yl)phenyl)-2-(piperidin-1-yl)acetamide(44)

2 mg of the desired compound 44 (28%) was obtained by the same processas step 5 of Example 1, using compound M-2.

¹H-NMR (CD₃OD, 400 MHz) δ 7.25 (d, J=7.8 Hz, 2H), 7.17-7.10 (m, 5H),7.05 (d, J=8.5 Hz, 2H), 6.86 (d, J=8.6 Hz, 2H), 6.79 (d, J=8.5 Hz, 2H),4.01 (s, 2H), 3.57 (m, 2H), 3.44 (t, J=6.7 Hz, 2H), 3.05 (m, 2H), 2.55(m, 2H), 1.90 (m, 6H), 1.56 (m, 2H). MS (ESI) m/z: 471 [M+H]⁺.

[Example 118] Preparation of(E)-4-(5-hydroxy-2-phenyl-1-(4-((2-(piperidin-1-yl)ethyl)amino)phenyl)pent-1-en-1-yl)phenol(45)

Compound 44 (10 mg, 0.02 mmol) was added to tetrahydrofuran (2 mL), thetemperature was lowered to 0° C., and 1 M lithium aluminum hydride(0.051 mL, 0.05 mmol) was added thereto. Heating was performed at 60° C.for 12 hour. Water and ethyl acetate were further added to the reactionsolution and an organic layer was extracted. The organic layer was driedwith anhydrous Na₂SO₄ and filtered. The solvent was distilled underreduced pressure to obtain a residue, which was purified using columnchromatography and then dissolved in methanol:dichloromethane (1:1), thetemperature was lowered to 0° C., a 1M aqueous HCl solution was slowlyadded thereto, and distillation under reduced pressure was performed,thereby obtaining 0.5 mg of the desired compound 45 (6%).

¹H-NMR (CD₃OD, 400 MHz) δ 7.15-7.07 (m, 5H), 7.01 (d, J=8.4 Hz, 2H),6.74 (m, 4H), 6.50 (d, J=8.4 Hz, 2H), 3.61 (m, 2H), 3.48 (m, 2H), 3.40(m, 6H), 2.51 (m, 2H), 1.81 (m, 4H), 1.55 (m, 4H). MS (ESI) m/z: 457[M+H]⁺.

[Example 119] Preparation of2-((3aR,6aS)-3a,6a-dimethylhexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-N-(4-((E)-5-hydroxy-1-(4-hydroxyphenyl)-2-phenylpent-1-en-1-yl)phenyl)acetamide(46)

Step 1: Preparation of tert-butyl(3aR,6aS)-5-(2-((4-((E)-5-methoxy-5-oxo-2-phenyl-1-(4-(pivaloyloxy)phenyl)pent-1-en-1-yl)phenyl)amino)-2-oxoethyl)-3a,6a-dimethylhexahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylate(N-1)

11 mg of the desired compound N-1 (99%) was obtained by the same processas step 1 of Example 106, using compound M-1.

Step 2: Preparation of tert-butyl(3aR,6aS)-5-(2-((4-((E)-5-hydroxy-1-(4-hydroxyphenyl)-2-phenylpent-1-en-1-yl)phenyl)amino)-2-oxoethyl)-3a,6a-dimethylhexahydropyrrolo[3,4-c]pyrrole-2(1H)-carboxylate(N-2)

4 mg of the desired compound N-2 (39%) was obtained by the same processas step 5 of Example 1, using compound N-1.

Step 3: Preparation of2-((3aR,6aS)-3a,6a-dimethylhexahydropyrrolo[3,4-c]pyrrol-2(1H)-yl)-N-(4-((E)-5-hydroxy-1-(4-hydroxyphenyl)-2-phenylpent-1-en-1-yl)phenyl)acetamide(46)

1 mg of the desired compound 46 (38%) was obtained by the same processas step 1 of Example 40, using compound N-2.

¹H-NMR (CD₃OD, 400 MHz) δ 7.23 (d, J=8.4 Hz, 2H), 7.17-7.08 (m, 5H),7.03 (d, J=8.4 Hz, 2H), 6.83 (d, J=8.5 Hz, 2H), 6.77 (d, J=8.4 Hz, 2H),4.24 (s, 2H), 3.43 (m, 8H), 2.52 (m, 2H), 1.52 (m, 4H). MS (ESI) m/z:498 [M+H]⁺.

[Example 120] Preparation of(Z)-1-(4-(5-hydroxy-1-(4-(4-isopropylpiperazin-1-yl)phenyl)-2-phenylpent-1-en-1-yl)phenyl)guanidine(50)

Step 1: Preparation of(Z)-5-(4-aminophenyl)-5-(4-(4-isopropylpiperazin-1-yl)phenyl)-4-phenylpent-4-en-1-ol(0-1)

Compound 26b (5 mg, 11 μmol), N,N′-di-boc-thiourea (3 mg, 0.01 mmol),mercury (II) chloride (3 mg, 0.01 mmol), and triethylamine (5 μL, 0.03mmol) were added to dimethylformamide (1 mL), and heated at roomtemperature for 12 hours. The solvent was distilled under reducedpressure to obtain a residue, which was purified using columnchromatography, thereby obtaining 6 mg of the desired compound 0-1(84%).

Step 2: Preparation of(Z)-1-(4-(5-hydroxy-1-(4-(4-isopropylpiperazin-1-yl)phenyl)-2-phenylpent-1-en-1-yl)phenyl)guanidine(50)

0.5 mg of the desired compound 50 (9%) was obtained by the same processas step 1 of Example 40, using compound O-1.

MS (ESI) m/z: 498 [M+H]⁺.

[Example 121] Preparation of(E)-4-(4-(5-hydroxy-1-(4-hydroxyphenyl)-2-phenylpent-1-en-1-yl)phenyl)piperazine-1-carboximidamide(52)

Step 1: Preparation of tert-butyl((E)-((tert-butoxycarbonyl)imino)(4-(4-((E)-5-hydroxy-1-(4-hydroxyphenyl)-2-phenylpent-1-en-1-yl)phenyl)piperazin-1-yl)methyl)carbamate(P-1)

5 mg of the desired compound P-1 (36%) was obtained by the same processas step 1 of Example 120, using compound 20c.

Step 2: Preparation of(E)-4-(4-(5-hydroxy-1-(4-hydroxyphenyl)-2-phenylpent-1-en-1-yl)phenyl)piperazine-1-carboximidamide(52)

0.7 mg of the desired compound 52 (23%) was obtained by the same processas step 1 of Example 40, using compound P-1.

¹H-NMR (CD₃OD, 400 MHz) δ 7.16-7.07 (m, 5H), 7.01 (m, 2H), 6.78 (m, 2H),6.67 (m, 2H), 6.40 (d, J=8.6 Hz, 2H), 3.67 (m, 2H), 3.28 (m, 2H), 3.48(m, 1H), 3.41 (t, J=6.7 Hz, 2H), 3.19 (m, 2H), 3.13 (m, 1H), 2.51 (m,2H), 1.54 (m, 2H). MS (ESI) m/z: 457 [M+H]⁺.

[Experimental Example 1] ERRγ, ERRα, ERRβ, ERα binding assay

1) ERRγ Binding Assay (Inverse Agonist Assay)

The arylethene derivative of the present invention was sequentiallyadded to a 384 well plate from a concentration of 10 μM to a finalconcentration of two-fold dilution. Then, a GST-bound ERR gammaligand-binding domain (LBD) was added to a final concentration of 5 nM,and a fluorescein-conjugated coactivator PGC1a and a Tb-a-GST antibodywere added to 500 nM and 5 nM, respectively. After all reagents wereadded, a reaction was carried out with gentle shaking at 20° C. for 1hour, and after the reaction, a binding activity was measured by aTR-FRET method. That is, excitation at 340 nm was performed, eachemission value at 495 nm and 520 nm was measured, the result assay was avalue measured at 490 nm/a value measured at 520 nm, and an analysisprogram was Prism 6.

2) ERRα/ERRβ/ERα Binding Assay (Selectivity Test)

In an ERR alpha binding assay, GST-bound ERR alpha LBD was used, and allexperiments other than that was the same as the ERR gamma binding assay.

In an ERR beta binding assay, GST-bound ERR alpha LBD was used so that afinal concentration was 10 nM and a fluorescein-conjugated coactivatorPGC1a was 250 nM, and all experiments other than that was the same asthe ERR gamma binding assay.

In an ER alpha binding assay, a GST-bound ER alpha ligand-binding domain(LBD) was added to a 384 well plate to which the arylethene derivativeof the present invention was added to a final concentration of 7.3 nM.Then, a fluorescein-conjugated coactivator PGC1a and a Tb-a-GST antibodywere added to 250 nM and 5 nM, respectively, and beta-estradiol as anagonist was added to a final concentration of 4 nM. All subsequentexperiments was the same as the ERR gamma binding assay.

The results of Experiment Example 1 are shown in the following Table 12.

TABLE 12 Binding Assay, IC50 (μM) Example Cmpd No. ERRγ ERRα ERRβ ERα 4 6d 0.093 >10 0.262 5  6e 0.067 >10 0.408 6  6f 0.059 >10 0.296 7  6g0.030 >10 0.331 8  6h 0.042 >10 1.1 10  6j 0.034 11  6k 0.026 16 13b0.064 >10 3.928 >10 20 18a 0.040 >10 1.33 1.24 21 18b 0.017 >100.861 >10 23 18d 0.060 >10 1.7 24 18e 0.060 >10 2.8 >10 25 18f 0.026 >101.017 >10 27 18h 0.032 >10 0.99 >10 28 18i 0.050 >10 3.04 1.142 29 18j0.025 >10 1.045 >10 30 18k 0.035 >10 >10 1.943 33 18n 0.029 8.25 1.098.8 40 20a 0.028 8.65 0.98 >10 41 20b 0.027 >10 2.097 >10 49 20j0.048 >10 4.93 0.992 58 22g 0.095 >10 6.766 4.371 60 22i 0.056 >10 1.6652.093 62 22k 0.049 >10 4.822 1.381 64 22m 0.078 >10 7.818 >10 67 22p0.099 9.7 1.648 2.116 68 22q 0.053 >10 2.002 1.658 71 22t 0.096 >103.564 5.94 87 27a 0.026 4.318 0.07 0.413 106 38a 0.079 >10 8.13 0.437107 38b 0.050 >10 3.07 0.256 108 38c 0.091 >10 >10 0.408 109 38d0.083 >10 8.8 0.276 GSK5182 0.107 >10 >10 2

[Experimental Example 2] ERRγ Inverse Agonist Functional Assay

AD293 was cultured in a 24-well plate for 24 hours, using a DMEM Highglucose culture medium (Hyclone, USA) to which 0.5% FBS was added at aconcentration of 9×10⁴/well. The culture medium was replaced with a DMEMHigh glucose culture medium to which 10% FBS was added, treatment wasperformed with a mixture of a TransIT-LT1 transfection reagent (Mirus,USA) and pCMX-Gal4-ERRγ, pFR-luciferase reporter plasmid, pCMV-β-gal,and culturing was performed for 24 hours. Thereafter, a luciferaseactivity assay and a β-gal assay were performed, respectively, with alysate obtained after treatment with the arylethene derivative of thepresent invention for 24 hours. All results were derived from three ormore independent repetitive experiments.

The results are shown in the following Table 13, in which “Cpds” refersto an inverse agonist functional activity when the compound was treated,“Ref 5182” refers to an activity of a reference compound GSK5182 fordata verification for every essay, and “Cpds/Ref 5182” refers to anactivity degree of the arylethene derivative of the present invention,relative to the reference compound.

TABLE 13 Functional Assay at 10 μM (% of control) Example Cmpd No. CpdsRef 5182 Cpds/Ref 5182 2  6b 7.97 3.08 2.59 3  6c 4.64 3.08 1.51 4  6d5.2 3.08 1.69 5  6e 5.61 3.08 1.82 6  6f 6.75 3.08 2.19 7  6g 4.94 3.081.60 8  6h 5.49 3.08 1.78 9  6i 5 3.08 1.62 10  6j 2.44 3.15 0.77 11  6k2.5 3.15 0.79 14  7a 2.71 3.15 0.86 16 13b 1.5 3.15 0.48 17 13c 10.583.15 3.36 20 18a 4.83 4.09 1.18 21 18b 2.45 3.15 0.78 22 18c 1.31 3.150.42 23 18d 9.65 9.97 0.97 24 18e 2.6 2.93 0.89 25 18f 2.57 3.15 0.82 2618g 1.62 1.93 0.84 27 18h 4.47 4.09 1.09 28 18i 1.88 1.93 0.97 29 18j3.28 2.72 1.21 30 18k 1.08 0.95 1.14 32 18m 2.97 2.93 1.01 33 18n 3.364.09 0.82 34 18o 23.2 9.97 2.33 38 18s 2.92 2.93 1.00 19 18t 11.1 9.971.11 39 18u 5.33 0.95 5.61 40 20a 3.15 4.09 0.77 41 20b 3.58 4.09 0.8842 20c 2.72 2.93 0.93 43 20d 3.32 4.09 0.81 44 20e 10.8 9.97 1.08 45 20f5.76 2.93 1.97 47 20h 3.57 3.15 1.13 49 20j 1.41 1.02 1.38 52 22a 3.14.09 0.76 53 22b 2.94 4.09 0.72 55 22d 1.96 3.15 0.62 56 22e 1.93 3.150.61 58 22g 1.84 1.93 0.95 59 22h 0.6 0.93 0.65 60 22i 3.37 2.72 1.24 6222k 0.77 0.93 0.83 64 22m 0.75 0.93 0.81 65 22n 3.47 2.72 1.28 69 22r0.64 0.95 0.67 70 22s 6.33 5.97 71 22t 6.19 5.97 72 22u 5.12 5.97 73 22v5.51 5.97 80 22ac 0.61 0.93 0.66 82 22ae 0.67 0.93 0.72 83 26a 2.66 3.150.84 85 26c 4.56 0.95 4.80 87 27a 2.1 2.72 0.77 95 28a 1.14 0.95 1.20 9828d 1.69 0.95 1.78 99 28e 4.76 4.09 1.16 100 28f 4 4.09 0.98 101 28g 5.14.09 1.25 102 30a 2.94 0.95 3.09 106 38a 1.45 1.02 1.42 107 38b 1.271.02 1.25 108 38c 4.06 4.09 0.99 109 38d 3.16 4.09 0.77 110 39 3.38 1.023.31 111 40 2.8 2.93 0.96 112 42a 4.37 2.93 1.49 113 42b 11.6 9.97 1.16114 42c 2.6 2.93 0.89 115 42d 11.7 2.93 3.99 116 42e 26.04 4.09 6.37 11744 12.94 4.09 3.16 118 45 1.58 1.02 1.55 119 46 78.46 3.15 24.91 GSK51823~10

[Experimental Example 3] In Vitro Absorption, Distribution, Metabolism,Excretion, and Toxicity (ADME)/Tox Evaluation

1) Cytochrome P450 (CYP450) Activity Inhibition Evaluation

Human liver microsomes (0.25 mg/ml) with 0.1 M phosphate buffer solution(pH 7.4), a substrate drug cocktail of five drug metabolizing enzymes(Phenacetin 50 μM, Diclofenac 10 μM, S-mephenytoin 100 μM,Dextromethorphan 5 μM, Midazolam 2.5 μM), and the arylethene derivativeof the present invention were added at concentrations of 0 μM and 10 μM,respectively, culturing was performed at 37° C. for 5 minutes inadvance, a NADPH generation system solution was added, and culturing wasperformed at 37° C. for 15 minutes. Thereafter, in order to complete thereaction, an acetonitrile solution containing an internal standardmaterial (Terfenadine) was added thereto, centrifugation (14,000 rpm, 4°C.) was performed for 5 minutes, and a supernatant was injected into aLC-MS/MS system to analyze the metabolites of the substrate drug,thereby evaluating drug metabolism enzyme inhibition by the arylethenederivative of the present invention.

The results are shown in the following Table 14.

TABLE 14 Cmpd CYP inhibition (% of control) Example No. 1A2 2C9 2C19 2D63A4 3  6c 91.9 77.9 81.6 96.4 81.3 5  6e 79.6 51.6 64.1 80.6 68.1 7  6g97.8 75.4 81.8 81.7 83.7 8  6h 103 62.8 83.2 90.7 75.4 9  6i 98.1 86.782.6 93.6 79.4 10  6j 98.7 85.6 89.1 85.2 74.6 18 13d >100 97 89.4 >10090.2 20 18a 88 86.1 81.2 63.5 66.4 21 18b 89.8 73.4 69.7 68.9 53 22 18c92.2 81.7 80 75.6 64.1 23 18d 88 67.7 81.2 98.4 64 24 18e >100 91.384.6 >100 84 25 18f >100 62.3 72.8 68.2 70.9 27 18h 93.5 96.9 86.3 71.564.5 29 18j >100 87 74 86.6 77.6 38 18s >100 81.8 82.4 >100 77.9 19 18t100 83 93 98 106 40 20a 85 76 61.9 70.7 55 49 20j 93.6 77.1 62 89.3 66.250 20k 79.3 87.6 78.9 96.2 83.6 53 22b 100.4 98.4 96.6 93.5 90.8 5522d >100 >100 >100 82.5 66.1 56 22e 94.9 61.3 58.4 71.8 58.2 59 22h >10097.9 >100 69.1 72.2 61 22j 95.7 88 89.6 97.1 85 62 22k >100 97 >100 73.774.7 63 22l 96.2 88.3 97 99 80.7 64 22m 97 80.3 >100 70.7 58.1 65 22n 9997.4 83 82.6 88.3 67 22p 88.1 81.8 61.5 79.6 90.6 68 22q 99.9 77.7 92.877.6 81 69 22r >100 >100 >100 91.8 82.7 80 22ac >100 >100 >100 86.2 8582 22ae 59.4 >100 >100 68.4 81.5 83 26a >100 53.2 89 85.5 75.4 84 26b99.2 92.5 98.7 81.9 60.9 86 26d >100 96.6 >100 86.1 57.3 108 38c 79.255.3 58.8 54.3 61.5 109 38d 79.2 55.3 58.8 54.3 57.6 110 39 82.5 74.473.9 67.8 59 GSK5182 84.6 72.9 78.2 82.3 83

2) Microsomal Stability Evaluation

Four liver microsomes (Human, Dog, Rat, Mouse 0.5 mg/ml) with a 0.1 Mphosphate buffer solution (pH 7.4), and the arylethene derivative of thepresent invention were added to a concentration of 1 μM, culturing wasperformed at 37° C. for 5 minutes in advance, a NADPH regenerationsystem solution was added thereto, and culturing was performed at 37° C.for 30 minutes. Thereafter, in order to complete the reaction, anacetonitrile solution containing an internal standard material(chlorpropamide) was added thereto, centrifugation (14,000 rpm, 4° C.)was performed for 5 minutes, and a supernatant was injected into aLC-MS/MS system to analyze the substrate drug, thereby evaluatingmetabolism stability to the arylethene derivative of the presentinvention.

The results are shown in the following Table 15.

TABLE 15 MS(Microsomal Stability) (%) Example Cmpd No. human dog ratmouse 3  6c 128.0 45.2 14  7a 49.0 64.7 59.5 32.1 16 13b 61.2 66.2 1813d 67.3 59.9 21 18b 54.2 40.0 19.7 22 18c 96.9 72.8 25 18f 79.2 65.747.6 26 18g 87.9 84.9 85.3 84.1 27 18h >100 64.4 74.2 48.5 29 18j 67.958.0 62.8 54.9 30 18k 65.4 61.2 54.9 26.9 19 18t 66.9 80.5 40 20a 65.184.7 41 20b 44 58 71.5 28.8 42 20c 55.8 53.7 43 20d 83 54 11.3 44 20e78.1 82.1 45 20f 83.9 99.5 49 20j 76.8 68.3 84.5 25.3 50 20k 93.6 84.989.0 81.3 53 22b 63.0 28.1 42.5 6.4 55 22d 57.6 60.2 57 22f 97.0 78.180.0 66.3 58 22g 42.8 54.3 43.2 10.7 59 22h 46.9 22.4 43.8 16.2 60 22i36.0 91.5 92.6 83.6 61 22j 65.2 54.9 65.5 34.9 62 22k 59.7 59.6 69.023.4 63 22l 71.8 67.9 53.4 25.8 64 22m 55.3 55.0 53.5 24.2 65 22n 56.372.0 47.8 37.8 67 22p 72.3 77.7 75.3 40.4 68 22q 63.7 43.8 67.6 17.6 6922r 62.0 56.9 49.9 24.5 70 22s 34.9 35.4 75.6 21.7 72 22u 40.0 10.0 44.317.2 82 22ae 48.4 33.7 32.2 21.5 83 26a 72.3 46.7 84 26b 49.1 40.6 31.414.3 86 26d 77.5 51.5 72.2 38.3 87 27a 66.2 50.3 65.0 32.7 89 27c 83.581.2 79.4 42.1 90 27d 63.7 43.8 67.6 17.6 99 28e 56.4 59.6 107 38b 41.841.8 53.4 38.4 109 38d 6.9 11.7 11.8 110 39 71.1 88.4 69.9 63.1 111 4064.7 72.0 GSK5182 42.8-45.1 9.6 26.0-29.1 6.8

3) Parallel Artificial Membrane Permeability Assay (PAMPA) Evaluation

PAMPA is a method which has been developed for testing cell membranepermeability of a material in a test tube, and has been performed usinga lipid tri-layer PVDF membrane available from Corning Gentest (NY, US),the used reagents were all purchased from Sigma (MO, US). First, a testmaterial is diluted in PBS (pH 7.4) to a final concentration of 10 mM,300 mL of the solution is added to the bottom well of a 96-transwellequipped with a PVDF membrane, and 200 mL of PBS is added to the upperwell. Then, a plate is reacted at 25° C. for 5 hours, 20 mL of thesolution in each well is transferred to a new container, and 80 mL ofacetonitrile containing an internal standard material (4 mMchloropropamide) is added thereto. A concentration of the material inthe solution is analyzed using LC-MS/MS (ThermoFisher Scientific, MO,US), and the transmittance of the material is calculated according tothe equation reported in the reference document.

Reference document: A novel design of artificial membrane for improvingthe PAMPA model. Chen X, Murawski A, et al. Pharmaceutical Research.25:1511, 2007

The results are shown in the following Table 16.

TABLE 16 Permeability Example Cmpd No. Pampa (10⁻⁶ cm/s) 11  6k 0.12 1613b 0.93 18 13d 0.14 20 18a 0.46 21 18b 2.09 22 18c 5.73 23 18d 0.14 2418e 1.11 25 18f 4.84 26 18g 4.8 27 18h 0.37 28 18i 1.16 29 18j 1.61 3018k 1.29 33 18n 0.63 38 18s 0.18 40 20a 0.52 41 20b 3.61 43 20d 0.23 4520f 0.14 55 22d 0.38 56 22e 6.16 57 22f 3.96 60 22i 0.35 67 22p 0.2 6822q 0.77 83 26a 1.58 86 26d 1.04 87 27a 2.22 89 27c 0.68 90 27d 0.34 11342b 0.1 GSK5182 0.11~0.82

4) hERG Channel Binding Inhibition Evaluation

An E-4031 (effective IC50: 10-90 nM) compound as a positive control wasdiluted stepwise with 3-fold, a pre-prepared membrane containing a hERGchannel and a fluorescent tracer were mixed and reacted for 4 hours, andthen a polarization values for each concentration were measured toobtain IC₅₀. For the arylethene derivative of the present invention,fluorescence intensity (excitation at 530 nm, emission at 590 nm) at aconcentration of stepwise diluted 16 points was measured and comparedwith a DMSO solvent control.

A hERG fluorescence polarization assay (Invitrogen: PV5365) kit wasused.

The results are shown in the following Table 17.

TABLE 17 Example Cmpd No. hERG IC50 (μM) 4  6d >30 6  6f >30 7  6g 5.4 8 6h 18.0 9  6i >30 20 18a 18.7 21 18b >30 22 18c 11.6 23 18d >20 25 18f18.9 26 18g 17.9 27 18h 15.3 28 18i >30 29 18j 17.0 30 18k 24.6 41 20b7.1 43 20d 15.0 49 20j 12.7 50 20k 26.1 55 22d 6.0 56 22e 20.7 57 22f5.8 58 22g 22.0 60 22i >30 61 22j 8.8 65 22n 14.6 68 22q 10.9 69 22r16.4 70 22s 10.6 72 22u 6.1 73 22v 5.8 83 26a >30 86 26d 26.0 106 38a16.1 110 39 9.5 GSK5182 >30

[Experimental Example 4] In Vivo Pharmacokinetics (In Vivo PK)Evaluation

In order to investigate pharmacokinetic behavior when intravenously ororally administrating the compound of the present invention to a rat,rats weighing at least 200 g were used to perform the followingexperiment, and the results are shown in the following Table 18.

A. Experimental Method

1. An oral administration group fasts the day before.

2. Blood of each animal is collected at 0 hour.

3. Into a tail vein of an intravenous administration group (IV), a drugis injected at a dose of 1 mg/kg (syringe).

4. To an oral administration group (PO), a drug is orally administeredat a dose of 10 mg/kg (oral zondec)

5. After administration, blood of the intravenous administration groupwas collected through a jugular vein 8 times at 0.08, 0.25, 0.5, 1, 2,4, 6, and 8 hours. One collected blood amount is 400 to 500 ul.

6. After administration, blood of the oral administration group wascollected through a jugular vein 6 times at 0.25, 0.5, 1, 4, 6 and 8hours. One collected blood amount is 400 to 500 ul.

7. Each blood is mixed with a 3.8% sodium citrate solution and stored onice.

8. Supernatant plasma is collected by a centrifuge.

9. The supernatant plasma was injected into a LC-MS/MS system and thedrug is analyzed.

TABLE 18 Cmpd Administration AUC_(all) AUC_(INF) BA C_(max)Cl(observed)/F T_(max) t_(1/2) Vss Example No. group (μMh) (μMh) (%)(μM) (mL/min/kg) (h) (h) (L/Kg) GSK5182 IV 0.89 0.94 44 2.3 9.1 PO 0.680.78 8.4 0.13 134 1.9 20 18a IV 0.42 0.49 75 3.8 25.2 PO 0.81 1.29 21.40.21 277 1.1 21 18b IV 0.58 0.64 58 3.1 9.8 PO 0.31 0.36 8.7 0.12 10650.6 24 18e IV 0.06 0.06 546 0.8 39.1 PO 0.07 0.09 11.5 0.01 430 2.8 2518f IV 0.28 0.29 139 2.5 30.3 PO 2.93 7.00 41.2 0.45 2.4 29 18j IV 0.550.60 59 3.2 9.8 PO 1.07 1.61 19.6 0.21 3 30 18k IV 0.99 1.00 37 1.2 3.0PO 4.18 4.71 42.4 0.97 1.7 2.4 44 20e IV 0.28 0.34 139 4.4 52.8 PO — — —— 60 22i IV 0.49 0.54 69 3.0 11.7 PO 2.24 4.52 45.3 0.38 2.2 64 22m IV0.34 0.00 110 2.4 19.9 PO 0.73 0.00 24.3 0.13 2.4 4.3 69 22r IV 0.850.87 45 1.8 4.6 PO 4.93 5.29 58.2 1.51 1 2.3 70 22s IV 0.44 81.1 2.213.2 PO 0.09 0.11 18.5 0.02 2855 1.3

[Experimental Example 5] Experiment on Anaplastic Thyroid Cancer

1. Materials and Method

1.1. Cells

CAL-62 which is an anaplastic thyroid cancer cell line was purchasedfrom Deutsche Sammlung von Mikroorganismen and Zellkulturen. The celllines were all maintained in a DMEM medium highly supplemented with 10%FBS, 1% antibiotic-antifungal agent (Hyclone), at 37° C. under theatmosphere of 5% CO₂. A retrovirus from which an enhanced fireflyluciferase gene (effluc) is expressed was treated with CAL-62 cells toestablish cell lines in which the effluc genes are stably expressed. Thethus-established cell lines were referred to as CAL-62/effluc cells.

1.2. ¹²⁵I Uptake Assay

The cells were plated in a 24-well plate for 24 hours, treated withcompound 18a, produced into a 100 mM stock solution in DMSO, and storedat −80° C. for 24 hours. After adsorbing a drug-containing medium, thecells were washed with 1 mL of HBSS, and incubated with 500μ of a Hank'balanced salt solution (HBSS) containing 0.5% bovine serum albumin(bHBSS), 3.7 kBq carrier-free ¹²⁵I (Perkin-Elmer), and 10 μmol/L ofsodium iodide (inactive 740 MBq/mmol) at 37° C. for 30 minutes.Thereafter, the cells were washed twice with ice-cold bHBSS, and lysedwith 500 μl of 2% sodium dodecyl sulfate (SDS). Radioactivity wasmeasured using a gamma counter (Cobra II; Canberra Packard, PackardBioscience). The radioactivity of the cells was normalized using a totalprotein concentration determined by a BCA kit (Pierce Protein Biology).

1.3. ¹²⁵I Uptake Assay Depending on Compound 18a Drug Concentration

The cells were treated with compound 18a at various concentrations(vehicle, 6, 12 uM), and then a ¹²⁵I uptake test was performed asdescribed above.

1.4. ¹²⁵I Uptake Inhibition Assay by KClO₄

The cells were pre-incubated with 300 μM KClO₄ (as a specific inhibitorto NIS) for 30 minutes to inhibit iodine uptake, and then a ¹²⁵I uptaketest was performed as described above.

1.5. ¹²⁵I uptake inhibition assay by MAK kinase inhibitor The cells werepre-incubated with PD98059 or U0126 (as a specific inhibitor to MAPkinase) for 30 minutes to inhibit iodine uptake, and then a 125I uptaketest was performed as described above.

1.6 Quantitative RT-PCR

Total RNA was separated using Trizol (Invitrogen, Carlsbad, Calif.).Total RNA (2 ug) was reverse-transcribed into cDNA with RevertAid FirstStrand cDNA Synthesis Kit (Thermo Scientific, Pittsburgh, Pa.). Geneswere amplified with a ViiA 7 Real-Time PCR System instrument (AppliedBiosystems) using the primer of each target gene and YBR Green PCRmaster mix (Applied Biosystems, Foster City, Calif.), using a cDNAtemplate. The primer sequence of each target gene is as follows: ERRγ(forward, 5′-CAG ACG CCA GTG GGA GCT A-3′; reverse, 5′-TGG CGA GTC AAGTCC GTT CT-3′), NIS (forward, 5′-TCT AAC CGA TGC TCA CCT CTT CTG-3′;reverse, 5′-AGA TGA TGG CAC CTC CTT GAA CC-3′), and acidic ribosomalprotein 36B4 (forward, 5′-CCA CGC TGC TGA ACA TGC T-3′; reverse, 5′-TCGAAC ACC TGC TGG ATG AC-3′). Each target gene was normalized using a 36B4gene.

1.7. Clonogenic Assay

The cells were plated in a 6-well plate, and allowed to stand for 24hours. The cells were treated with 12 μM compound 18a for 24 hours, thedrug-containing medium was discarded, and the cells were washed twicewith PBS. Thereafter, the medium was replaced with DMEM for 6 hours inthe presence or absence of 50 μCi ¹³¹I (KIRAMS, Korea). The cells werewashed with cold bHBSS, and allowed to stand in a normalized culturemedium for a time corresponding to six doublings. Finally, the cellswere fixed in a 4% paraformaldehyde (PFA) solution and stained with0.05% crystal violet. Control colonies having more than 50 cells and¹³¹I-treated colonies were counted.

1.8. Western Blot

The cells were treated with or without compound 18a for 24 hours, washedtwice with cold PBS, and lysed with a RIPA (Roche) buffer containing acomplete protease inhibitor cocktail. In the case of cell membraneprotein for NIS, samples were prepared using a protein biotinylation kit(EZ-Link™ Sulfo-NHS-Biotin, Thermo Scientific) according to themanufacturer's instruction. Briefly, any one of non-treated cells ortreated cells were washed twice with ice-cold PBS/CM (PBS containing 0.1mM calcium chloride and 1 mM magnesium chloride, pH 7.3), and incubatedwith EZ link NHS-sulfo-SS-biotin in PBS/CM (1 mg/mL) at 4° C. for 30minutes. The reaction was quenched by washing twice using cold 100 mMglycine in PBS/CM, and further incubated with 100 mM glycine in PBS/CMat 4° C. for 20 minutes. Thereafter, the cells were constantly shaken at4° C. for 1 hour, and rapidly washed twice using PBS/CM before beinglysed using a RIPA buffer (Roche) containing a protease inhibitorcocktail and a phosphatase inhibitor. The lysate was centrifuged at16,000 g, at 4° C. for 30 minutes. A portion of the supernatant was usedfor a total cell protein immune blot. The remaining sample was incubatedwith 100 μL streptavidin beads (Thermo Scientific) at room temperaturefor 1 hour to be used for obtaining membrane protein. The beads werewashed three times using a RIPA buffer, the bound protein was elutedusing 50 μL of Laemmli buffer (62.5M Tris, pH 6.8; 20% glycerol; 2% SDS;5% b-mercaptoethanol; and 0.01% bromophenol blue) at room temperaturefor 30 minutes. Equivalent amounts of the total cell membrane proteinand biotinylated cell membrane protein were loaded on each lane, andresolved by a Bis-Tris gel (Invitrogen) with a 4-12% slope. The proteinwas moved to a 0.2 μm PVDF membrane (Invitrogen). The membrane wasincubated with a primary rat monoclonal human NIS-specific antibody(dilution 1:1000, Thermo Scientific, Catalog #: MS-1653-P1, clone:FP5A), and then incubated with a HRP-conjugated secondary antibody atroom temperature. ECL-Plus (Amersham Pharmacia) was used for detecting aperoxidase activity, depending on the manufacturer's method. Similarly,even in the case of other protein, an equivalent amount of protein wasloaded to each lane, and resolved by a Bis-Tris gel (Invitrogen) with a4-12% slope. The protein was moved to a 0.2 μm PVDF membrane(Invitrogen). The membrane was incubated with a primary antibody (ERRγ,pERK1/2, β-actin) at 4° C. for one night, and then incubated with anappropriate HRP-conjugated secondary antibody at room temperature.According to the manufacturer's protocol, the peroxidase activity wasdetected using ECL-Plus. A band density was determined using an ImageJsoftware.

1.9. Animal Experiment

Nude mice (Balb/c nu/nu, female, 6 weeks old) were used, and all animalswere normally raised in DMRC center animal laboratory of KyungbukNational University Hospital in Chilgok. 5×10⁶ CAL-62/effluc cells weresubcutaneously injected into the left femoral region of the nude mouseto form a tumor. The tumor was extracted, divided into small pieces (20mg or more), and then intradermally injected into the nude mouse to forma tumor.

After forming the tumor, the CAL-62/effluc mouse tumor model was dividedinto the following groups: Group 1: vehicle, Group 2: 100 mpk compound18a, Group 3: 100 mpk compound 18a. To the mice of each group, thevehicle (100% PEG) and compound 18a (100 mpk, 200 mpk) were orallyadministered daily for 6 days. In order to observe a difference in tumorgrowth between before administration and after administration, opticalimaging (bioluminescent imaging) was performed. While the drug wasadministered, a weight change of the mouse was observed every other day.

In addition, in order to confirm a change in a 125I uptake increase inthe CAL-62/effluc tumor, an organ distribution study (Bio-distributionstudy) was performed as follows. After finally administrating the drug,¹²⁵I (5 uCi/mouse) was administered to the mouse by intravenousinjection on the next day. After 4 hours of administration, all organsincluding a parent tumor were extracted, and each organ was weighed.Thereafter, each organ was transferred to a 5 mL test tube, andradioactivity in the organ was measured using a gamma counter. A ¹²⁵Iuptake degree in the organ was expressed by percentage injected dose pergram (% ID/g).

1.10. Animal Image

For obtaining an optical image, D-luciferin (3 mg/mouse) wasintraperitoneally injected to the mouse. After about 10 minutes ofinjection, the mouse was anesthetized by inhalation (I-2% isofluranegas), and then positioned on a IVIS Lumina III (PerkinElmer) imagingbed. The time for obtaining the image was automatically set, and then anoptical image was obtained. A Living imaging software (version 2.12,PerkinElmer) was used to quantify an optical image signal from thetumor.

1.11. Statistical Analysis

All data was represented as an average ±, and statistical significancewas determined using a Student test of GraphPad Prism 5. A P value <0.05was regarded as being statistically significant.

2. Results

2.1 Increased Radioactive Iodine Uptake in ATC Cells by Compound 18a

After treatment with compound 18a, a significant increase of radioactiveiodine uptake in CAL62 cells was confirmed for each concentration andeach time (FIGS. 1 and 2). A maximum increase of iodine uptake wasobserved at a concentration of 12 uM of compound 18a. In order to testwhether the increased radioactive iodine uptake is related to regulationof a NIS function by compound 18a, KClO₄ which is an inhibitor specificto NIS was co-incubated with compound 18a-treated CAL62 cells, and achange in a radioactive iodine uptake level was observed. KClO₄completely blocks radioactive iodine uptake which was increased in thecompound 18a-treated cells (FIG. 3), which implies that the increasediodine uptake is directly related to the improved functional activity ofNIS mediated by compound 18a.

2.2 Endogenous ERRγ and NIS mRNA Expression Regulation by Compound 18ain ATC Cells

In order to determine the effect of compound 18a on an ERRγ mRNA levelin ATC cells, real-time PCR was performed using ERRγ- and NIS-specificprimers. As a result of treatment with compound 18a, it was confirmedthat ERRγ mRNA expression in CAL62 cells were significantly decreased(FIG. 4), and when compared with the vehicle treated group, theexpression was decreased by about 16 times. However, it was confirmedthat NIS mRNA expression was increased by about 2 times when comparedwith the vehicle-treated group.

2.3 Endogenous ERRγ Protein Regulation by Compound 18a in ATC Cells

In order to determine the effect of compound 18a on an ERRγ proteinlevel in ATC cells, immune blotting assay was performed usingERRγ-specific antibody. As a result of treatment with compound 18a, itwas confirmed that ERRγ protein expression in CAL62 cells weresignificantly decreased (FIG. 6), and when compared with the vehicletreated group, the expression was decreased by about 2.8 times (FIG. 7).

2.4 Increase of Membrane-Localized NIS Protein in ATC Cells ThroughActivation of Endogenous MPA Kinase Signaling by Compound 18a in ATCCells

A significant increase in a phosphorylated MPP kinase level such as p44and p42 ERK was found in ATC cells treated with compound 18a (FIG. 8).The relative increase of the phosphorylated forms oERK1 and ERK2f was2.2 times and 2.8 times, respectively (FIG. 9).

The radioactive iodine uptake increase (FIGS. 10 and 11) and therelative increase of the phosphorylated form of ERK1 and ERK2 bycompound 18a were completely inhibited by selective MEK inhibitors,PD98059 and U0126 (FIGS. 12 and 13).

In order to determine the effect of compound 18a on an ERRγ proteinlevel in ATC cells, immune blotting assay was performed usingNIC-specific antibody. As a result of treatment with compound 18a, itwas confirmed that total NIS protein (fully or partially glycosylatedform) expression in CAL62 cells were significantly increased (FIGS. 14and 15), and when compared with the vehicle treated group, theexpression was decreased by about 1.9 times. In order to determine theeffect of compound 18a on the state of NIS membrane protein, a change ina level of membranous total NIS protein collected from compound18a-treated CAL 62 cells using a cell membrane biotinylated kit wasexamined using an immune blotting examination using an NIS-specificantibody. Compound 18a derived a sharp increase in cellmembrane-localized NIS protein having mature and immature forms in ATCcells, as compared with control cells (FIG. 14). Qualitative analysis ofband intensity showed increases in membrane fully glycosylated andpartially glycosylated NIS protein in CAL62 cells by 8.1 times and 6.4times, respectively (FIG. 15).

2.5 Modification of I-131 Mediated Cytotoxicity by Compound 18a in ATCCells

A clone formation assay using I-131 showed a minimal cytotoxic effect inCAL62 cells treated with any one of compounds 18a and I-131 alone (FIG.16). Relative colony formability of I-131 or GSK5182 group was 92.9±5.8%and 94.5±10.8%, respectively in CAL62 cells (FIG. 17). However, as aresult of combining ¹³¹I and GSK5182, colony-formability wassignificantly decreased to about 58.5±7.4% in CAL-62 (FIG. 17).

2.6 Increase of Radioactive Iodine Uptake by Administration of Compound18a in ATC Tumor Model

The CAL62-effluc mouse tumor model was divided into the following groups(FIG. 18, Group 1: vehicle, Group 2: 100 mpk compound 18a, Group 3: 200mpk compound 18a). To the mice of each group, the vehicle (100% PEG) andcompound 18a (100 mpk, 200 mpk) were orally administered daily for 6days. In order to observe a difference in tumor growth between beforeadministration and after administration, optical imaging (bioluminescentimaging) was performed. After finally administering the drug, aradioactive isotope (I-125) was administered to the mice on the nextday, and after 2 hours, the mice were sacrificed, all organs thereofwere extracted, and a radiation level was measured with a gamma counter.It was confirmed that radioactive iodine uptake in CAL62 tumor wasconcentration-dependently increased by treatment with compound 18a (FIG.19). When compared with the vehicle group, the radioactive uptake wasincreased by 4.4 times and 16.2 times in the 100 mpk and 200 mpkcompound 18a groups, respectively. When observing the difference intumor growth using optical imaging, significant tumor growth inhibitoryefficiency was shown in the compound 18a group (FIG. 20). Drugconcentration-dependent tumor growth inhibitory efficiency was shown(FIG. 21). An abrupt weight change in the mice was not shown in allgroups (FIG. 22).

Hereinabove, although the present invention has been described in detailwith reference to the exemplary embodiments, it will be apparent tothose skilled in the art that various modifications and alterations maybe made without departing from the scope and spirit of the presentinvention. It should be understood that these modifications andalterations fall within the scope defined by the following claims.

INDUSTRIAL APPLICABILITY

The arylethene derivative of the present invention is a novel compound,and exhibits very high inhibitory activity to ERRγ as compared with aconventional GSK5182 compound, and at the same time, shows an effect ofimproved drug stability, pharmacological activity and toxicity. Thus,the arylethene derivative may be useful as efficient prophylactic agentand therapeutic agent for diseases mediated by ERRγ, in particular,metabolic diseases such as obesity, diabetes, hyperlipidemia, fattyliver, or atherosclerosis, as well as retinopathy, without side effects.

In addition, the arylethene derivative of the present invention mayspecifically and significantly inhibit ERRγ transcriptional activity ascompared with GSK5182, and as a result, cause a radioactive isotopeuptake increase from a cellular level to an animal level. Accordingly,the arylethene derivative of the present invention may significantlyincrease a treatment effect of radioactive iodine therapy for treatingcancer, and when administered to cancer cells, may effectively producecancer cells having an improved sodium iodide symporter (NIS) function,thereby having an excellent effect of being more easily applied torelated research and clinical practice for treating anaplastic thyroidcancer.

The invention claimed is:
 1. A compound represented by the followingChemical Formulae 2 to 5, or a stereoisomer, or pharmaceuticallyacceptable salt thereof:

wherein

denotes a single bond or a double bond; R¹ is (C3-C20)heterocycloalkyl,(C3-C20)heteroaryl, —O—(CH₂)_(m)R¹¹, —(CH₂)_(m)—R¹², —NH—(CH₂)_(m)—R¹³,—NHCO—(CH₂)_(n)—R¹⁴, or —SiR¹⁶R¹⁷—(CH₂)_(m)—R¹⁵; R¹¹ is selected fromthe following structures:

wherein R³¹ and R³² are independently of each other hydrogen,(C1-C10)alkyl, (C3-C10)cycloalkyl, (C2-C10)alkenyl, amidino,(C1-C10)alkoxycarbonyl, hydroxy(C1-C10)alkyl, ordi(C1-C10)alkylamino(C1-C10)alkyl; and L is S; R¹² to R¹⁵ areindependently of one another (C3-C20)heterocycloalkyl; R¹⁶ and R¹⁷ areindependently of each other (C1-C20)alkyl; m is an integer of 1 to 3;and n is an integer of 0 or 1; Ar is (C6-C20)aryl or (C3-C20)heteroaryl,in which the aryl or heteroaryl of Ar may be further substituted by oneor more selected from the group consisting of hydroxy, halogen,(C1-C20)alkyl, halo(C1-C20)alkyl, (C1-C20)alkoxy, nitro, cyano,—NR²¹R²², (C1-C20)alkylcarbonyloxy, (C1-C20)alkylcarbonylamino,guanidino, —SO₂—R²³ and —OSO₂—R²⁴; R²¹ and R²² are independently of eachother hydrogen, (C1-C20)alkylsulfonyl, or (C3-C20)cycloalkylsulfonyl;R²³ and R²⁴ are independently of each other (C1-C20)alkyl,halo(C1-C20)alkyl, or (C3-C20)cycloalkyl; R² is hydroxy, fluoro,(C1-C20)alkylcarbonyloxy, or (C1-C20)alkylsulfonyloxy; theheterocycloalkyl or heteroaryl of R¹ and the heterocycloalkyl of R¹² toR¹⁵ may be further substituted by one or more selected from the groupconsisting of (C1-C20)alkyl, (C3-C20)cycloalkyl, (C2-C20)alkenyl,amidino, (C1-C20)alkoxycarbonyl, hydroxy, hydroxy(C1-C20)alkyl, anddi(C1-C20)alkylamino(C1-C20)alkyl; the heterocycloalkyl is a monovalentradical of a non-aromatic heterocycle containing 1 to 4 heteroatomsselected from the group consisting of N, O and S, and is a saturated orunsaturated mono-, bi-, or spirocycle having a carbon atom or nitrogenatom in a ring as a binding site; and the heteroaryl is a monovalentradical of a heteroaromatic ring which is an aryl group containing 1 to4 heteroatoms selected from the group consisting of N, O and S as anaromatic ring backbone atom, and carbons as remaining aromatic ringbackbone atoms.
 2. The compound, or stereoisomer, or pharmaceuticallyacceptable salt thereof, of claim 1, wherein R¹ is(C3-C10)heterocycloalkyl, (C3-C10)heteroaryl, —O—(CH₂)_(m)—R¹¹,—(CH₂)_(m)—R¹², —NH—(CH₂)_(m)—R¹³, —NHCO—(CH₂)_(n)—R¹⁴, or—SiR¹⁶R¹⁷—(CH₂)_(m)—R¹¹; R¹² to R¹⁵ are independently of one another(C3-C10)heterocycloalkyl; R¹⁶ and R¹⁷ are independently of each other(C1-C10)alkyl; m is an integer of 1 to 3; n is an integer of 0 or 1; Aris (C6-C12)aryl or (C3-C12)heteroaryl, in which the aryl or heteroarylof Ar may be further substituted by one or more selected from the groupconsisting of hydroxy, halogen, (C1-C10)alkyl, halo(C1-C10)alkyl,(C1-C10)alkoxy, nitro, cyano, amino, (C1-C10)alkylsulfonylamino,(C3-C10)cycloalkylsulfonylamino, di((C1-C10)alkylsulfonyl)amino,(C1-C10)alkylcarbonyloxy, (C1-C10)alkylcarbonylamino, guanidino,(C1-C10)alkylsulfonyl, (C1-C10)alkylsulfonyloxy,halo(C1-C10)alkylsulfonyloxy, and (C3-C10)cycloalkylsulfonyloxy; R² ishydroxy, fluoro, (C1-C10)alkylcarbonyloxy, or (C1-C10)alkylsulfonyloxy;and the heterocycloalkyl or heteroaryl of R¹ and the heterocycloalkyl ofR¹² to R¹⁵ may be further substituted by one or more selected from thegroup consisting of (C1-C10)alkyl, (C3-C10)cycloalkyl, (C2-C10)alkenyl,amidino, (C1-C10)alkoxycarbonyl, hydroxy(C1-C10)alkyl, anddi(C1-C10)alkylamino(C1-C10)alkyl.
 3. The compound, or stereoisomer, orpharmaceutically acceptable salt thereof, of claim 2, wherein R¹ is(C3-C10)heterocycloalkyl or —O—(CH₂)_(m)—R¹¹; m is an integer of 1 to 3;and the heterocycloalkyl of R¹ may be further substituted by one or moreselected from the group consisting of (C1-C10)alkyl, (C3-C10)cycloalkyl,(C2-C10)alkenyl, amidino, (C1-C10)alkoxycarbonyl, hydroxy(C1-C10)alkyl,and di(C1-C10)alkylamino(C1-C10)alkyl.
 4. The compound, or stereoisomer,or pharmaceutically acceptable salt thereof, of claim 1, wherein theheterocycloalkyl of R¹ and R¹² to R¹⁵ is independently of each otherselected from the following structures:

wherein R³¹ and R³² are independently of each other hydrogen,(C1-C10)alkyl, (C3-C10)cycloalkyl, (C2-C10)alkenyl, amidino,(C1-C10)alkoxycarbonyl, hydroxy(C1-C10)alkyl, ordi(C1-C10)alkylamino(C1-C10)alkyl; and L is O or S.
 5. The compound, orstereoisomer, or pharmaceutically acceptable salt thereof, of claim 1,wherein the compound is represented by the following Chemical Formula 6:

wherein R¹ is (C3-C10)heterocycloalkyl or —O—(CH₂)_(m)—R¹¹; m is aninteger of 1 to 3; the heterocycloalkyl of R may be further substitutedby one or more selected from the group consisting of (C1-C10)alkyl,(C3-C10)cycloalkyl, (C2-C10)alkenyl, amidino, (C1-C10)alkoxycarbonyl,hydroxy(C1-C10)alkyl, and di(C1-C20)alkylamino(C1-C20)alkyl; Ar is(C6-C12)aryl or (C3-C12)heteroaryl, in which the aryl or heteroaryl ofAr may be further substituted by one or more selected from the groupconsisting of hydroxy, halogen, (C1-C10)alkyl, halo(C1-C10)alkyl,(C1-C10)alkoxy, nitro, cyano, amino, (C1-C10)alkylsulfonylamino,(C3-C10)cycloalkylsulfonylamino, di((C1-C10)alkylsulfonyl)amino,(C1-C10)alkylcarbonyloxy, (C1-C10)alkylcarbonylamino, guanidino,(C1-C10)alkylsulfonyl, (C1-C10)alkylsulfonyloxy,halo(C1-C10)alkylsulfonyloxy, and (C3-C10)cycloalkylsulfonyloxy; and R²is hydroxy, fluoro, (C1-C10)alkylcarbonyloxy, or(C1-C10)alkylsulfonyloxy.
 6. The compound, or stereoisomer, orpharmaceutically acceptable salt thereof, of claim 5, wherein R² ishydroxy; and R is heterocycloalkyl selected from the followingstructures:

wherein R³¹ and R³² are independently of each other hydrogen,(C1-C10)alkyl, (C3-C10)cycloalkyl, (C2-C10)alkenyl, amidino,(C1-C10)alkoxycarbonyl, hydroxy(C1-C10)alkyl, ordi(C1-C10)alkylamino(C1-C10)alkyl; and L is O or S.
 7. The compound, orstereoisomer, or pharmaceutically acceptable salt thereof, of claim 5,wherein R² is hydroxy; R¹ is —O—(CH₂)_(m)—R¹¹; m is an integer of 1 or2.
 8. The compound, or stereoisomer, or pharmaceutically acceptable saltthereof, of claim 1, wherein the compound is selected from the followingstructures:


9. The compound, or stereoisomer, or pharmaceutically acceptable saltthereof, of claim 5, wherein the compound is selected from the followingstructures: